Compositions and methods for inhibiting expression of a gene from the Ebola virus

ABSTRACT

The invention relates to a double-stranded ribonucleic acid (dsRNA) for inhibiting the expression of a gene from the Ebola virus.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.13/627,508, filed Sep. 26, 2012, which is a divisional of U.S. patentapplication Ser. No. 13/093,681, filed Apr. 25, 2011, issued as U.S.Pat. No. 8,354,390, which is a divisional of U.S. patent applicationSer. No. 12/789,414, filed May 27, 2010, issued as U.S. Pat. No.7,973,020, which is a divisional of U.S. patent application Ser. No.12/055,295, filed Mar. 25, 2008, issued as U.S. Pat. No. 7,759,320,which claims priority to U.S. Provisional Application Ser. No.60/908,793, filed Mar. 29, 2007, the entire contents of which are herebyincorporated by reference.

GOVERNMENT SUPPORT

This invention was made with government support under contract numberHHSN266200600012C, awarded by the Department of Health and HumanServices (DHHS). The government has certain rights in the invention.

REFERENCE TO A SEQUENCE LISTING

This application includes a Sequence Listing submitted electronically asa text file named 26560_US_Sequence₁₃ Listing.txt, created on May 15,2014, with a size of 757 kb and comprising 1108 sequences. The sequencelisting is hereby incorporated by reference.

FIELD OF THE INVENTION

This invention relates to double-stranded ribonucleic acid (dsRNA), andits use in mediating RNA interference to inhibit the expression of oneof the genes of the Ebola virus and the use of the dsRNA to treatpathological processes mediated by Ebola infection, such as systemichemorrhage and multi-organ failure.

BACKGROUND OF THE INVENTION

Ebola Virus

Minus-strand (−) RNA viruses are major causes of human suffering thatcause epidemics of serious human illness. In humans the diseases causedby these viruses include influenza (Orthomyxoviridae), mumps, measles,upper and lower respiratory tract disease (Paramyxoviridae), rabies(Rhabdoviridae), hemorrhagic fever (Filoviridae, Bunyaviridae andArenaviridae), encephalitis (Bunyaviridae) and neurological illness(Bomaviridae). Virtually the entire human population is thought to beinfected by many of these viruses.

The Ebola virus comes from the Filoviridae family, similar to theMarburg virus. It is named after the Ebola River in Zaire, Africa, nearwhere the first outbreak was noted by Dr. Ngoy Mushola in 1976 after asignificant outbreaks in both Yambuku, Zaire (now the DemocraticRepublic of the Congo), and Nzara, in western Sudan. Of 602 identifiedcases, there were 397 deaths.

The two strains identified in 1976 were named Ebola-Zaire (EBO-Z) andEbola-Sudan (EBO-S). The outbreak in Sudan showed a lower fatalityrate—50%—compared to the 90% mortality rate of the Zaire strain. In1990, a second, similar virus was identified in Reston, Va. amongstmonkeys imported from the Philippines, and was named Ebola-Reston.

Further outbreaks have occurred in Zaire/Congo (1995 and 2003), Gabon(1994, 1995 and 1996), and in Uganda (2000). A new subtype wasidentified from a single human case in the Côte d'Ivoire in 1994,EBO-CI.

Of around 1500 identified human Ebola infections, two-thirds of thepatients have died. The animal (or other) reservoir which sustains thevirus between outbreaks has not been identified.

Among humans, the Ebola virus is transmitted by direct contact withinfected body fluids such as blood.

The incubation period of Ebola hemorrhagic fever varies from two days tofour weeks. Symptoms are variable too, but the onset is usually suddenand characterised by high fever, prostration, myalgia, arthralgia,abdominal pains and headache. These symptoms progress to vomiting,diarrhea, oropharyngeal lesions, conjunctivitis, organ damage (notablythe kidney and liver) by co-localized necrosis, proteinuria, andbleeding both internal and external, commonly through thegastrointestinal tract. Death or recovery to convalescence occurs withinsix to ten days of onset of symptomology.

The development of a successful therapeutic for Ebola virus is along-sought and seemingly difficult endeavor. Although they cause only afew hundred deaths worldwide each year, filoviruses are considered asignificant world health threat and have many of the characteristicscommonly associated with biological weapons since they can be grown inlarge quantities, can be fairly stable, are highly infectious as anaerosol, and are exceptionally deadly. Filoviruses are relatively simpleviruses of 19 Kb genomes and consist of seven genes which encodenucleoprotein (NP), glycoprotein (GP), four smaller viral proteins(VP24, VP30, VP35 and VP40), and the RNA-dependent RNA polymerase (Lprotein) all in a single strand of negative-sensed RNA. Administrationof type I interferons, therapeutic vaccines, immune globulins,ribavirin, and other nucleoside analogues have been somewhat successfulin rodent Ebola virus models, but not in nonhuman primate infectionmodels.

In view of the severity of the diseases caused by (−) RNA viruses, inparticular members of the Filoviridae family of viruses, and the lack ofeffective prevention or therapies, it is therefore an object of thepresent invention to provide therapeutic compounds and methods fortreating a host infected with a (−) RNA virus.

siRNA

Double-stranded RNA molecules (dsRNA) have been shown to block geneexpression in a highly conserved regulatory mechanism known as RNAinterference (RNAi). WO 99/32619 (Fire et al.) discloses the use of adsRNA of at least 25 nucleotides in length to inhibit the expression ofgenes in C. elegans. dsRNA has also been shown to degrade target RNA inother organisms, including plants (see, e.g., WO 99/53050, Waterhouse etal.; and WO 99/61631, Heifetz et al.), Drosophila (see, e.g., Yang, D.,et al., Curr. Biol. (2000) 10:1191-1200), and mammals (see WO 00/44895,Limmer; and DE 101 00 586.5, Kreutzer et al.). This natural mechanismhas now become the focus for the development of a new class ofpharmaceutical agents for treating disorders that are caused by theaberrant or unwanted regulation of a gene.

Recent reports have indicated that in vitro, RNAi may show somepromising in reducing Ebola replication and providing protection inguinea pigs (Geisbert, et al., The Journal of Infectious Diseases, 193(2006), 1650-1657). However, the RNAi agents examined were not designedagainst all known Ebola strains and were not selected for stability andother properties needed for in vivo therapeutic RNAi agents.Accordingly, despite significant advances in the field of RNAi, thereremains a need for an agent that can selectively and efficiently silencea gene in the Ebola virus using the cell's own RNAi machinery that hasboth high biological activity and in vivo stability, and that caneffectively inhibit replication of the Ebola virus for use in treatingpathological processes mediated by Ebola infection.

SUMMARY OF THE INVENTION

The invention provides double-stranded ribonucleic acid (dsRNA), as wellas compositions and methods for inhibiting the expression of the Ebolavirus in a cell or mammal using such dsRNA. The invention also providescompositions and methods for treating pathological conditions anddiseases caused by Ebola viral infection, such as systemic hemorrhageand multi-organ failure. The dsRNA featured in the invention includes anRNA strand (the antisense strand) having a region which is less than 30nucleotides in length, generally 19-24 nucleotides in length, and issubstantially complementary to at least part of an mRNA transcript of agene from the Ebola virus.

In one embodiment, the invention provides dsRNA molecules for inhibitingthe expression of a gene of the Ebola virus and viral replication. ThedsRNA comprises at least two sequences that are complementary to eachother. The dsRNA comprises a sense strand comprising a first sequenceand an antisense strand comprising a second sequence. The antisensestrand comprises a nucleotide sequence which is substantiallycomplementary to at least part of an mRNA encoded by a gene from theEbola virus, and the region of complementarity is less than 30nucleotides in length, generally 19-24 nucleotides in length. The dsRNA,upon contact with a cell infected with the Ebola virus, inhibits theexpression of a gene from the Ebola virus by at least 40%.

For example, the dsRNA molecules of the invention can include a firstsequence of the dsRNA that is selected from the group consisting of thesense sequences of Table 2 and the second sequence selected from thegroup consisting of the antisense sequences of Table 2. The dsRNAmolecules featured in the invention can include naturally occurringnucleotides or can include at least one modified nucleotide, such as a2′-O-methyl modified nucleotide, a nucleotide comprising a5′-phosphorothioate group, and a terminal nucleotide linked to acholesteryl derivative. Alternatively, the modified nucleotide may bechosen from the group of: a 2′-deoxy-2′-fluoro modified nucleotide, a2′-deoxy-modified nucleotide, a locked nucleotide, an a basicnucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide,morpholino nucleotide, a phosphoramidate, and a non-natural basecomprising nucleotide. Generally, such modified sequence will be basedon a first sequence of said dsRNA selected from the group consisting ofthe sense sequences of Table 2 and a second sequence selected from thegroup consisting of the antisense sequences of Table 2.

In another embodiment, the invention provides a cell having a dsRNA ofthe invention. The cell is generally a mammalian cell, such as a humancell.

In another embodiment, the invention provides a pharmaceuticalcomposition for inhibiting the replication of the Ebola virus in anorganism, generally a human subject. The composition includes one ormore of the dsRNA of the invention and a pharmaceutically acceptablecarrier or delivery vehicle.

In another embodiment, the invention provides a method for inhibitingthe expression of a gene in the Ebola virus in a cell, including thefollowing steps:

-   -   (a) introducing into the cell a double-stranded ribonucleic acid        (dsRNA), wherein the dsRNA includes at least two sequences that        are complementary to each other. The dsRNA includes a sense        strand having a first sequence and an antisense strand having a        second sequence. The antisense strand includes a region of        complementarity which is substantially complementary to at least        a part of an mRNA encoded by the Ebola virus, and wherein the        region of complementarity is less than 30 nucleotides in length,        generally 19-24 nucleotides in length, and optionally, wherein        the dsRNA, upon contact with a cell infected with the Ebola        virus, inhibits expression of a gene from the Ebola virus by at        least 40%, such as in an assay described herein (e.g., a        fluorscence-based assay); and    -   (b) maintaining the cell produced in step (a) for a time        sufficient to obtain degradation of the mRNA transcript of a        Ebola gene, thereby inhibiting expression of a gene from the        Ebola virus in the cell.

In another embodiment, the invention provides methods for treating,preventing or managing pathological processes mediated by Ebolainfection, such as systemic hemorrhage and multi-organ failure,comprising administering to a patient in need of such treatment,prevention or management a therapeutically or prophylactically effectiveamount of one or more of the dsRNAs of the invention.

In another embodiment, the invention provides vectors for inhibiting theexpression of a gene of the Ebola virus in a cell, comprising aregulatory sequence operably linked to a nucleotide sequence thatencodes at least one strand of a dsRNA of the invention.

In another embodiment, the invention provides a cell comprising a vectorfor inhibiting the expression of a gene of the Ebola virus in a cell.The vector comprises a regulatory sequence operably linked to anucleotide sequence that encodes at least one strand of a dsRNA of theinvention.

In one aspect, the invention provides for a method of increasing thelife-span of a subject (e.g., a mammal, such as a human or nonhumanprimate) infected with an Ebola virus. The method includes administeringa dsRNA to the subject, where the dsRNA includes an antisense RNA strandhaving a region which is less than 30 nucleotides in length, generally19-24 nucleotides in length, and is substantially complementary to atleast part of an mRNA transcript of a gene from the Ebola virus. ThedsRNA is administered in an amount sufficient to increase the lifespanof the subject. In one embodiment, the dsRNA includes an antisense RNAstrand having a region that is substantially complementary to at leastpart of an mRNA transcript of a gene selected from the VP30, VP35, NP,L, VP24, VP40 and GP genes. In a preferred embodiment, the dsRNAincludes an antisense RNA strand having a region that is substantiallycomplementary to at least part of an mRNA transcript of the VP35 gene.In some embodiments, the subject does not experience a decrease in oneor both of lymphocyte or platelet count after administration of thedsRNA. In other embodiments, the subject does not experience an increasein cytokine levels (e.g., IFN-alpha or TNF-alpha levels).

In another aspect, the invention features a method of decreasing viraltitre in a subject (e.g., a mammal, such as a human or nonhuman primate)infected with an Ebola virus. The method includes administering a dsRNAto the subject, where the dsRNA includes an antisense RNA strand havinga region which is less than 30 nucleotides in length, generally 19-24nucleotides in length, and is substantially complementary to at leastpart of an mRNA transcript of a gene from the Ebola virus. In oneembodiment, the dsRNA includes an antisense RNA strand having a regionthat is substantially complementary to at least part of an mRNAtranscript of the VP35 gene. In another embodiment, the subject does notexperience a decrease in one or both of lymphocyte or platelet countafter administration of the dsRNA. In other embodiments, the subjectdoes not experience an increase in cytokine levels (e.g., IFN-alpha orTNF-alpha levels).

In another aspect, the invention features a method of sustaininglymphocyte or platelet count in a mammal (e.g., a human or nonhumanprimate) infected with an Ebola virus. The method includes administeringa dsRNA to the subject, where the dsRNA includes an antisense RNA strandhaving a region which is less than 30 nucleotides in length, generally19-24 nucleotides in length, and is substantially complementary to atleast part of an mRNA transcript of a gene from the Ebola virus. In oneembodiment, the dsRNA includes an antisense RNA strand having a regionthat is substantially complementary to at least part of an mRNAtranscript of the VP35 gene. In other embodiments, the subject does notexperience an increase in cytokine levels (e.g., IFN-alpha or TNF-alphalevels).

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a graph showing that siRNAs formulated with lipidoid LNP01protected mice from a lethal Ebola virus challenge.

FIG. 2 is a graph showing that a single injection of a liposomallyformulated siRNA delivered by ip or iv protected mice from a lethalEbola challenge. VP35 siRNA was AD-11570

FIG. 3 is the structure of NP98 lipid.

FIG. 4 is a graph showing that siRNAs formulated with DODMA protectedmice from a lethal Ebola virus challenge.

FIG. 5 is a graph showing that siRNAs formulated with DODMA wereeffective down to 0.04 mg/kg to protect mice injected with Ebola.

FIG. 6 is a graph showing that siRNAs formulated with DODMA wereeffective to protect guinea pigs from a lethal Ebola virus challenge.

FIG. 7 is a graph showing the efficacy of siRNAs against different Ebolagenes formulated with DODMA in a guinea pig model of Ebola.

FIG. 8 is a graph presenting the observed decrease in viral titers inthe serum of mice following administration of LNP01-formulated VP35siRNA.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides double-stranded ribonucleic acid (dsRNA), as wellas compositions and methods for inhibiting the expression of a gene fromthe Ebola virus in a cell or mammal using the dsRNA. The invention alsoprovides compositions and methods for treating pathological conditionsand diseases in a mammal caused by Ebola infection using dsRNA. dsRNAdirects the sequence-specific degradation of mRNA through a processknown as RNA interference (RNAi).

The dsRNA of the invention comprises an RNA strand (the antisensestrand) having a region which is less than 30 nucleotides in length,generally 19-24 nucleotides in length, and is substantiallycomplementary to at least part of an mRNA transcript of a gene from theEbola virus. The use of these dsRNAs enables the targeted degradation ofmRNAs of genes that are implicated in replication and or maintenance ofEbola infection and the occurance of systemic hemorrhage and multi-organfailure in a subject infected with the Ebola virus. Using cell-based andanimal assays, the present inventors have demonstrated that very lowdosages of these dsRNA can specifically and efficiently mediate RNAi,resulting in significant inhibition of expression of a gene from theEbola virus. Thus, the methods and compositions of the inventioncomprising these dsRNAs are useful for treating pathological processesmediated by Ebolaviral infection by targeting a gene involved in Ebolarelication and/or maintainance in a cell.

The following detailed description discloses how to make and use thedsRNA and compositions containing dsRNA to inhibit the expression of agene from the Ebola virus, as well as compositions and methods fortreating diseases and disorders caused by the infection with the Ebolavirus, such as systemic hemorrhage and multi-organ failure. Thepharmaceutical compositions of the invention comprise a dsRNA having anantisense strand comprising a region of complementarity which is lessthan 30 nucleotides in length, generally 19-24 nucleotides in length,and is substantially complementary to at least part of an RNA transcriptof a gene from the Ebola virus, together with a pharmaceuticallyacceptable carrier.

Accordingly, certain aspects of the invention provide pharmaceuticalcompositions comprising the dsRNA of the invention together with apharmaceutically acceptable carrier, methods of using the compositionsto inhibit expression of a gene in a gene from the Ebola virus, andmethods of using the pharmaceutical compositions to treat diseasescaused by infection with the Ebola virus.

I. Definitions

For convenience, the meaning of certain terms and phrases used in thespecification, examples, and appended claims, are provided below. Ifthere is an apparent discrepancy between the usage of a term in otherparts of this specification and its definition provided in this section,the definition in this section shall prevail.

“G,” “C,” “A” and “U” each generally stand for a nucleotide thatcontains guanine, cytosine, adenine, and uracil as a base, respectively.However, it will be understood that the term “ribonucleotide” or“nucleotide” can also refer to a modified nucleotide, as furtherdetailed below, or a surrogate replacement moiety. The skilled person iswell aware that guanine, cytosine, adenine, and uracil may be replacedby other moieties without substantially altering the base pairingproperties of an oligonucleotide comprising a nucleotide bearing suchreplacement moiety. For example, without limitation, a nucleotidecomprising inosine as its base may base pair with nucleotides containingadenine, cytosine, or uracil. Hence, nucleotides containing uracil,guanine, or adenine may be replaced in the nucleotide sequences of theinvention by a nucleotide containing, for example, inosine. Sequencescomprising such replacement moieties are embodiments of the invention.

As used herein, “Ebola viruses”, are members of the family Filoviridae,are associated with outbreaks of highly lethal hemorrhagic fever inhumans and nonhuman primates. Human pathogens include Ebola Zaire, EbolaSudan, and Ebola Ivory Coast. Ebola Reston is a monkey pathogen and isnot considered a significant human pathogen. The natural reservoir ofthe virus is unknown and there are currently no available vaccines oreffective therapeutic treatments for filovirus infections. The genome ofEbola virus consists of a single strand of negative sense RNA that isapproximately 19 kb in length. This RNA contains seven sequentiallyarranged genes that produce 8 mRNAs upon infection. Ebola virions, likevirions of other filoviruses, contain seven proteins: a surfaceglycoprotein (GP), a nucleoprotein (NP), four virion structural proteins(VP40, VP35, VP30, and VP24), and an RNA-dependent RNA polymerase (L)(Feldmann et al. (1992) Virus Res. 24, 1-19; Sanchez et al., (1993)Virus Res. 29, 215-240; reviewed in Peters et al. (1996) In FieldsVirology, Third ed. pp. 1161-1176. Fields, B. N., Knipe, D. M., Howley,P. M., et al. eds. Lippincott-Raven Publishers, Philadelphia). Theglycoprotein of Ebola virus is unusual in that it is encoded in two openreading frames. Transcriptional editing is needed to express thetransmembrane form that is incorporated into the virion (Sanchez et al.(1996) Proc. Natl. Acad. Sci. USA 93, 3602-3607; Volchkov et al, (1995)Virology 214, 421-430).

As used herein, “target sequence” refers to a contiguous portion of thenucleotide sequence of an mRNA molecule formed during the transcriptionof a gene from the Ebola virus, including mRNA that is a product of RNAprocessing of a primary transcription product.

As used herein, the term “strand comprising a sequence” refers to anoligonucleotide comprising a chain of nucleotides that is described bythe sequence referred to using the standard nucleotide nomenclature.

As used herein, and unless otherwise indicated, the term“complementary,” when used to describe a first nucleotide sequence inrelation to a second nucleotide sequence, refers to the ability of anoligonucleotide or polynucleotide comprising the first nucleotidesequence to hybridize and form a duplex structure under certainconditions with an oligonucleotide or polynucleotide comprising thesecond nucleotide sequence, as will be understood by the skilled person.Such conditions can, for example, be stringent conditions, wherestringent conditions may include: 400 mM NaCl, 40 mM PIPES pH 6.4, 1 mMEDTA, 50° C. or 70° C. for 12-16 hours followed by washing. Otherconditions, such as physiologically relevant conditions as may beencountered inside an organism, can apply. The skilled person will beable to determine the set of conditions most appropriate for a test ofcomplementarity of two sequences in accordance with the ultimateapplication of the hybridized nucleotides.

This includes base-pairing of the oligonucleotide or polynucleotidecomprising the first nucleotide sequence to the oligonucleotide orpolynucleotide comprising the second nucleotide sequence over the entirelength of the first and second nucleotide sequence. Such sequences canbe referred to as “fully complementary” with respect to each otherherein. However, where a first sequence is referred to as “substantiallycomplementary” with respect to a second sequence herein, the twosequences can be fully complementary, or they may form one or more, butgenerally not more than 4, 3 or 2 mismatched base pairs uponhybridization, while retaining the ability to hybridize under theconditions most relevant to their ultimate application. However, wheretwo oligonucleotides are designed to form, upon hybridization, one ormore single stranded overhangs, such overhangs shall not be regarded asmismatches with regard to the determination of complementarity. Forexample, a dsRNA comprising one oligonucleotide 21 nucleotides in lengthand another oligonucleotide 23 nucleotides in length, wherein the longeroligonucleotide comprises a sequence of 21 nucleotides that is fullycomplementary to the shorter oligonucleotide, may yet be referred to as“fully complementary” for the purposes of the invention.

“Complementary” sequences, as used herein, may also include, or beformed entirely from, non-Watson-Crick base pairs and/or base pairsformed from non-natural and modified nucleotides, in as far as the aboverequirements with respect to their ability to hybridize are fulfilled.

The terms “complementary”, “fully complementary” and “substantiallycomplementary” herein may be used with respect to the base matchingbetween the sense strand and the antisense strand of a dsRNA, or betweenthe antisense strand of a dsRNA and a target sequence, as will beunderstood from the context of their use.

As used herein, a polynucleotide which is “substantially complementaryto at least part of” a messenger RNA (mRNA) refers to a polynucleotidewhich is substantially complementary to a contiguous portion of the mRNAof interest (e.g., encoding Ebola). For example, a polynucleotide iscomplementary to at least a part of a Ebola mRNA if the sequence issubstantially complementary to a non-interrupted portion of a mRNAencoding Ebola.

The term “double-stranded RNA” or “dsRNA,” as used herein, refers to acomplex of ribonucleic acid molecules, having a duplex structurecomprising two anti-parallel and substantially complementary, as definedabove, nucleic acid strands. The two strands forming the duplexstructure may be different portions of one larger RNA molecule, or theymay be separate RNA molecules. Where the two strands are part of onelarger molecule, and therefore are connected by an uninterrupted chainof nucleotides between the 3′-end of one strand and the 5′ end of therespective other strand forming the duplex structure, the connecting RNAchain is referred to as a “hairpin loop”. Where the two strands areconnected covalently by means other than an uninterrupted chain ofnucleotides between the 3′-end of one strand and the 5′ end of therespective other strand forming the duplex structure, the connectingstructure is referred to as a “linker”. The RNA strands may have thesame or a different number of nucleotides. The maximum number of basepairs is the number of nucleotides in the shortest strand of the dsRNAminus any overhangs that are present in the duplex. In addition to theduplex structure, a dsRNA may comprise one or more nucleotide overhangs.dsRNAs as used herein are also referred to as “siRNAs” (shortinterfering RNAs).

As used herein, a “nucleotide overhang” refers to the unpairednucleotide or nucleotides that protrude from the duplex structure of adsRNA when a 3′-end of one strand of the dsRNA extends beyond the 5′-endof the other strand, or vice versa. “Blunt” or “blunt end” means thatthere are no unpaired nucleotides at that end of the dsRNA, i.e., nonucleotide overhang. A “blunt ended” dsRNA is a dsRNA that isdouble-stranded over its entire length, i.e., no nucleotide overhang ateither end of the molecule.

The term “antisense strand” refers to the strand of a dsRNA whichincludes a region that is substantially complementary to a targetsequence. As used herein, the term “region of complementarity” refers tothe region on the antisense strand that is substantially complementaryto a sequence, for example a target sequence, as defined herein. Wherethe region of complementarity is not fully complementary to the targetsequence, the mismatches are most tolerated in the terminal regions and,if present, are generally in a terminal region or regions, e.g., within6, 5, 4, 3, or 2 nucleotides of the 5′ and/or 3′ terminus.

The term “sense strand,” as used herein, refers to the strand of a dsRNAthat includes a region that is substantially complementary to a regionof the antisense strand.

“Introducing into a cell”, when referring to a dsRNA, means facilitatinguptake or absorption into the cell, as is understood by those skilled inthe art. Absorption or uptake of dsRNA can occur through unaideddiffusive or active cellular processes, or by auxiliary agents ordevices. The meaning of this term is not limited to cells in vitro; adsRNA may also be “introduced into a cell”, wherein the cell is part ofa living organism. In such instance, introduction into the cell willinclude the delivery to the organism. For example, for in vivo delivery,dsRNA can be injected into a tissue site or administered systemically.In vitro introduction into a cell includes methods known in the art suchas electroporation and lipofection.

The terms “silence” and “inhibit the expression of”, in as far as theyrefer to a gene from the Ebola virus, herein refer to the at leastpartial suppression of the expression of a gene from the Ebola virus, asmanifested by a reduction of the amount of mRNA transcribed from a genefrom the Ebola virus which may be isolated from a first cell or group ofcells in which a gene from the Ebola virus is transcribed and which hasor have been treated such that the expression of a gene from the Ebolavirus is inhibited, as compared to a second cell or group of cellssubstantially identical to the first cell or group of cells but whichhas or have not been so treated (control cells). The degree ofinhibition is usually expressed in terms of

${\frac{\left( {{mRNA}\mspace{14mu}{in}\mspace{14mu}{control}\mspace{14mu}{cells}} \right) - \left( {{mRNA}\mspace{14mu}{in}\mspace{14mu}{treated}\mspace{14mu}{cells}} \right)}{\left( {{mRNA}\mspace{14mu}{in}\mspace{14mu}{control}\mspace{14mu}{cells}} \right)} \cdot 100}\%$

Alternatively, the degree of inhibition may be given in terms of areduction of a parameter that is functionally linked to Ebola genometranscription, e.g. the amount of protein encoded by a gene from theEbola virus, or the number of cells displaying a certain phenotype, e.ginfection with the Ebola virus. In principle, Ebola genome silencing maybe determined in any cell expressing the target, either constitutivelyor by genomic engineering, and by any appropriate assay. However, when areference is needed in order to determine whether a given dsRNA inhibitsthe expression of a gene from the Ebola virus by a certain degree andtherefore is encompassed by the instant invention, the assay provided inthe Examples below shall serve as such reference.

For example, in certain instances, expression of a gene from the Ebolavirus is suppressed by at least about 20%, 25%, 35%, or 50% byadministration of the double-stranded oligonucleotide of the invention.In some embodiment, a gene from the Ebola virus is suppressed by atleast about 60%, 70%, or 80% by administration of the double-strandedoligonucleotide of the invention. In some embodiments, a gene from theEbola virus is suppressed by at least about 85%, 90%, or 95% byadministration of the double-stranded oligonucleotide of the invention.

As used herein in the context of Ebola expression, the terms “treat”,“treatment”, and the like, refer to relief from or alleviation ofpathological processes mediated by Ebola infection. In the context ofthe present invention insofar as it relates to any of the otherconditions recited herein below (other than pathological processesmediated by Ebola expression), the terms “treat”, “treatment”, and thelike mean to relieve or alleviate at least one symptom associated withsuch condition, or to slow or reverse the progression of such condition,or to reduce the amount of virus present in the infected subject.

As used herein, the phrases “therapeutically effective amount” and“prophylactically effective amount” refer to an amount that provides atherapeutic benefit in the treatment, prevention, or management ofpathological processes mediated by Ebola infection or an overt symptomof pathological processes mediated by Ebola expression or the amountvirus present in the patient. The specific amount that istherapeutically effective can be readily determined by ordinary medicalpractitioner, and may vary depending on factors known in the art, suchas, e.g. the type of pathological processes mediated by Ebola infection,the patient's history and age, the stage of pathological processesmediated by Ebola infection, and the administration of otheranti-pathological processes mediated by Ebola infection.

As used herein, a “pharmaceutical composition” comprises apharmacologically effective amount of a dsRNA and a pharmaceuticallyacceptable carrier. As used herein, “pharmacologically effectiveamount,” “therapeutically effective amount” or simply “effective amount”refers to that amount of an RNA effective to produce the intendedpharmacological, therapeutic or preventive result. For example, if agiven clinical treatment is considered effective when there is at leasta 25% reduction in a measurable parameter associated with a disease ordisorder, a therapeutically effective amount of a drug for the treatmentof that disease or disorder is the amount necessary to effect at least a25% reduction in that parameter. Further, the pharmaceutical compositioncan be designed to enhance targeting cells involved in Ebola infectionsuch as dendritic cells, macrophages, hepatocytes, and other parenchymalcells.

The term “pharmaceutically acceptable carrier” refers to a carrier foradministration of a therapeutic agent. Such carriers include, but arenot limited to, saline, buffered saline, dextrose, water, glycerol,ethanol, and combinations thereof. The term specifically excludes cellculture medium. For drugs administered orally, pharmaceuticallyacceptable carriers include, but are not limited to pharmaceuticallyacceptable excipients such as inert diluents, disintegrating agents,binding agents, lubricating agents, sweetening agents, flavoring agents,coloring agents and preservatives. Suitable inert diluents includesodium and calcium carbonate, sodium and calcium phosphate, and lactose,while corn starch and alginic acid are suitable disintegrating agents.Binding agents may include starch and gelatin, while the lubricatingagent, if present, will generally be magnesium stearate, stearic acid ortalc. If desired, the tablets may be coated with a material such asglyceryl monostearate or glyceryl distearate, to delay absorption in thegastrointestinal tract.

As used herein, a “transformed cell” is a cell into which a vector hasbeen introduced from which a dsRNA molecule may be expressed.

II. Double-stranded Ribonucleic Acid (dsRNA)

In one embodiment, the invention provides double-stranded ribonucleicacid (dsRNA) molecules for inhibiting the expression of a gene from theEbola virus in a cell or mammal, wherein the dsRNA comprises anantisense strand comprising a region of complementarity which iscomplementary to at least a part of an mRNA formed in the expression ofa gene from the Ebola virus, and wherein the region of complementarityis less than 30 nucleotides in length, generally 19-24 nucleotides inlength, and wherein said dsRNA, upon contact with a cell expressing thegene from the Ebola virus, inhibits the expression of the Ebola virusgene by at least 40%.

The dsRNA comprises two RNA strands that are sufficiently complementaryto hybridize to form a duplex structure. One strand of the dsRNA (theantisense strand) comprises a region of complementarity that issubstantially complementary, and generally fully complementary, to atarget sequence, derived from the sequence of an mRNA formed during theexpression of a gene from the Ebola virus, the other strand (the sensestrand) comprises a region which is complementary to the antisensestrand, such that the two strands hybridize and form a duplex structurewhen combined under suitable conditions. Generally, the duplex structureis between 15 and 30, more generally between 18 and 25, yet moregenerally between 19 and 24, and most generally between 19 and 21 basepairs in length. Similarly, the region of complementarity to the targetsequence is between 15 and 30, more generally between 18 and 25, yetmore generally between 19 and 24, and most generally between 19 and 21nucleotides in length. The dsRNA of the invention may further compriseone or more single-stranded nucleotide overhang(s).

The dsRNA can be synthesized by standard methods known in the art asfurther discussed below, e.g., by use of an automated DNA synthesizer,such as are commercially available from, for example, Biosearch, AppliedBiosystems, Inc. In one embodiment, a gene from the Ebola virus is thefrom human Ebola genome. In specific embodiments, the antisense strandof the dsRNA comprises the sense sequences of Table 2 and the secondsequence is selected from the group consisting of the antisensesequences of Table 2. Alternative antisense agents that target elsewherein the target sequence provided in Table 2 can readily be determinedusing the target sequence and the flanking Ebola sequence.

In further embodiments, the dsRNA comprises at least one nucleotidesequence selected from the groups of sequences provided in Table 2. Inother embodiments, the dsRNA comprises at least two sequences selectedfrom this group, wherein one of the at least two sequences iscomplementary to another of the at least two sequences, and one of theat least two sequences is substantially complementary to a sequence ofan mRNA generated in the expression of a gene from the Ebola virus.Generally, the dsRNA comprises two oligonucleotides, wherein oneoligonucleotide is described as the sense strand in Table 2 and thesecond oligonucleotide is described as the antisense strand in Table 2

The skilled person is well aware that dsRNAs comprising a duplexstructure of between 20 and 23, but specifically 21, base pairs havebeen hailed as particularly effective in inducing RNA interference(Elbashir et al., EMBO 2001, 20:6877-6888). However, others have foundthat shorter or longer dsRNAs can be effective as well. In theembodiments described above, by virtue of the nature of theoligonucleotide sequences provided in Table 2, the dsRNAs of theinvention can comprise at least one strand of a length of minimally 21nucleotides. It can be reasonably expected that shorter dsRNAscomprising one of the sequences of Table 2 minus only a few nucleotideson one or both ends may be similarly effective as compared to the dsRNAsdescribed above. Hence, dsRNAs comprising a partial sequence of at least15, 16, 17, 18, 19, 20, or more contiguous nucleotides from one of thesequences of Table 2, and differing in their ability to inhibit theexpression of a gene from the Ebola virus in a FACS assay as describedherein below by not more than 5, 10, 15, 20, 25, or 30% inhibition froma dsRNA comprising the full sequence, are contemplated by the invention.Further dsRNAs that cleave within the target sequence provided in Table2 can readily be made using the Ebola virus sequence and the targetsequence provided.

In addition, the RNAi agents provided in Table 2 identify a site in theEbola virus mRNA that is susceptible to RNAi based cleavage. As such thepresent invention further includes RNAi agents that target within thesequence targeted by one of the agents of the present invention. As usedherein a second RNAi agent is said to target within the sequence of afirst RNAi agent if the second RNAi agent cleaves the message anywherewithin the mRNA that is complementary to the antisense strand of thefirst RNAi agent. Such a second agent will generally consist of at least15 contiguous nucleotides from one of the sequences provided in Table 2coupled to additional nucleotide sequences taken from the regioncontiguous to the selected sequence in a gene from the Ebola virus. Forexample, the last 15 nucleotides of SEQ ID NO:1 combined with the next 6nucleotides from the target Ebola genome produces a single strand agentof 21 nucleotides that is based on one of the sequences provided inTable 2.

The dsRNA of the invention can contain one or more mismatches to thetarget sequence. In one embodiment, the dsRNA of the invention containsno more than 3 mismatches. If the antisense strand of the dsRNA containsmismatches to a target sequence, it is preferable that the area ofmismatch not be located in the center of the region of complementarity.If the antisense strand of the dsRNA contains mismatches to the targetsequence, it is preferable that the mismatch be restricted to 5nucleotides from either end, for example 5, 4, 3, 2, or 1 nucleotidefrom either the 5′ or 3′ end of the region of complementarity. Forexample, for a 23 nucleotide dsRNA strand which is complementary to aregion of a gene from the Ebola virus, the dsRNA generally does notcontain any mismatch within the central 13 nucleotides. The methodsdescribed within the invention can be used to determine whether a dsRNAcontaining a mismatch to a target sequence is effective in inhibitingthe expression of a gene from the Ebola virus. Consideration of theefficacy of dsRNAs with mismatches in inhibiting expression of a genefrom the Ebola virus is important, especially if the particular regionof complementarity in a gene from the Ebola virus is known to havepolymorphic sequence variation within the population.

In one embodiment, at least one end of the dsRNA has a single-strandednucleotide overhang of 1 to 4, generally 1 or 2 nucleotides. dsRNAshaving at least one nucleotide overhang have unexpectedly superiorinhibitory properties than their blunt-ended counterparts. Moreover, thepresent inventors have discovered that the presence of only onenucleotide overhang strengthens the interference activity of the dsRNA,without affecting its overall stability. dsRNA having only one overhanghas proven particularly stable and effective in vivo, as well as in avariety of cells, cell culture mediums, blood, and serum. Generally, thesingle-stranded overhang is located at the 3′-terminal end of theantisense strand or, alternatively, at the 3′-terminal end of the sensestrand. The dsRNA may also have a blunt end, generally located at the5′-end of the antisense strand. Such dsRNAs have improved stability andinhibitory activity, thus allowing administration at low dosages, i.e.,less than 5 mg/kg body weight of the recipient per day. Generally, theantisense strand of the dsRNA has a nucleotide overhang at the 3′-end,and the 5′-end is blunt. In another embodiment, one or more of thenucleotides in the overhang is replaced with a nucleoside thiophosphate.

In yet another embodiment, the dsRNA is chemically modified to enhancestability. The nucleic acids of the invention may be synthesized and/ormodified by methods well established in the art, such as those describedin “Current protocols in nucleic acid chemistry”, Beaucage, S. L. et al.(Edrs.), John Wiley & Sons, Inc., New York, N.Y., USA, which is herebyincorporated herein by reference. Specific examples of dsRNA compoundsuseful in this invention include dsRNAs containing modified backbones orno natural internucleoside linkages. As defined in this specification,dsRNAs having modified backbones include those that retain a phosphorusatom in the backbone and those that do not have a phosphorus atom in thebackbone. For the purposes of this specification, and as sometimesreferenced in the art, modified dsRNAs that do not have a phosphorusatom in their internucleoside backbone can also be considered to beoligonucleosides.

Preferred modified dsRNA backbones include, for example,phosphorothioates, chiral phosphorothioates, phosphorodithioates,phosphotriesters, aminoalkylphosphotriesters, methyl and other alkylphosphonates including 3′-alkylene phosphonates and chiral phosphonates,phosphinates, phosphoramidates including 3′-amino phosphoramidate andaminoalkylphosphoramidates, thionophosphoramidates,thionoalkylphosphonates, thionoalkylphosphotriesters, andboranophosphates having normal 3′-5′ linkages, 2′-5′ linked analogs ofthese, and those) having inverted polarity wherein the adjacent pairs ofnucleoside units are linked 3′-5′ to 5′-3′ or 2′-5′ to 5′-2′. Varioussalts, mixed salts and free acid forms are also included.

Representative U.S. patents that teach the preparation of the abovephosphorus-containing linkages include, but are not limited to, U.S.Pat. Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,195;5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131;5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925;5,519,126; 5,536,821; 5,541,316; 5,550,111; 5,563,253; 5,571,799;5,587,361; and 5,625,050, each of which is herein incorporated byreference

Preferred modified dsRNA backbones that do not include a phosphorus atomtherein have backbones that are formed by short chain alkyl orcycloalkyl internucleoside linkages, mixed heteroatoms and alkyl orcycloalkyl internucleoside linkages, or ore or more short chainheteroatomic or heterocyclic internucleoside linkages. These includethose having morpholino linkages (formed in part from the sugar portionof a nucleoside); siloxane backbones; sulfide, sulfoxide and sulfonebackbones; formacetyl and thioformacetyl backbones; methylene formacetyland thioformacetyl backbones; alkene containing backbones; sulfamatebackbones; methyleneimino and methylenehydrazino backbones; sulfonateand sulfonamide backbones; amide backbones; and others having mixed N,O, S and CH2 component parts.

Representative U.S. patents that teach the preparation of the aboveoligonucleosides include, but are not limited to, U.S. Pat. Nos.5,034,506; 5,166,315; 5,185,444; 5,214,134; 5,216,141; 5,235,033;5,64,562; 5,264,564; 5,405,938; 5,434,257; 5,466,677; 5,470,967;5,489,677; 5,541,307; 5,561,225; 5,596,086; 5,602,240; 5,608,046;5,610,289; 5,618,704; 5,623,070; 5,663,312; 5,633,360; 5,677,437; and,5,677,439, each of which is herein incorporated by reference.

In other certain dsRNA mimetics, both the sugar and the internucleosidelinkage, i.e., the backbone, of the nucleotide units are replaced withnovel groups. The base units are maintained for hybridization with anappropriate nucleic acid target compound. One such oligomeric compound,an dsRNA mimetic that has been shown to have excellent hybridizationproperties, is referred to as a peptide nucleic acid (PNA). In PNAcompounds, the sugar backbone of an dsRNA is replaced with an amidecontaining backbone, in particular an aminoethylglycine backbone. Thenucleobases are retained and are bound directly or indirectly to azanitrogen atoms of the amide portion of the backbone. Representative U.S.patents that teach the preparation of PNA compounds include, but are notlimited to, U.S. Pat. Nos. 5,539,082; 5,714,331; and 5,719,262, each ofwhich is herein incorporated by reference. Further teaching of PNAcompounds can be found in Nielsen et al., Science, 1991, 254, 1497-1500.

Other embodiments featured in the invention include dsRNAs withphosphorothioate backbones and oligonucleosides with heteroatombackbones, and in particular—CH.sub.2—NH—CH.sub.2-,—CH.sub.2-N(CH.sub.3)—O—CH.sub.2-[known as a methylene(methylimino) orMMI backbone], —CH.sub.2-O—N(CH.sub.3)—CH.sub.2-,—CH.sub.2-N(CH.sub.3)—N(CH.sub.3)—CH.sub.2- and—N(CH.sub.3)—CH.sub.2-CH.sub.2- [wherein the native phosphodiesterbackbone is represented as —O—P—O—CH.sub.2-] of the above-referencedU.S. Pat. No. 5,489,677, and the amide backbones of the above-referencedU.S. Pat. No. 5,602,240. Also preferred are dsRNAs having morpholinobackbone structures of the above-referenced U.S. Pat. No. 5,034,506.

Modified dsRNAs may also contain one or more substituted sugar moieties.In some embodiment, the dsRNAs include one of the following at the 2′position: OH; F; O—, S—, or N-alkyl; O—, S—, or N-alkenyl; O—, S— orN-alkynyl; or O-alkyl-O-alkyl, wherein the alkyl, alkenyl and alkynylmay be substituted or unsubstituted C.sub.1 to C.sub.10 alkyl or C.sub.2to C.sub.10 alkenyl and alkynyl. Particularly preferred areO[(CH.sub.2).sub.nO].sub.mCH.sub.3, O(CH.sub.2).sub.nOCH.sub.3,O(CH.sub.2).sub.nNH.sub.2, O(CH.sub.2).sub.nCH.sub.3,O(CH.sub.2).sub.nONH.sub.2, andO(CH.sub.2).sub.nON[(CH.sub.2).sub.nCH.su- b.3)].sub.2, where n and mare from 1 to about 10. Other preferred dsRNAs comprise one of thefollowing at the 2′ position: C.sub.1 to C.sub.10 lower alkyl,substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl, SH,SCH.sub.3, OCN, Cl, Br, CN, CF.sub.3, OCF.sub.3, SOCH.sub.3,SO.sub.2CH.sub.3, ONO.sub.2, NO.sub.2, N.sub.3, NH.sub.2,heterocycloalkyl, heterocycloalkaryl, aminoalkylamino, polyalkylamino,substituted silyl, an RNA cleaving group, a reporter group, anintercalator, a group for improving the pharmacokinetic properties of andsRNA, or a group for improving the pharmacodynamic properties of andsRNA, and other substituents having similar properties. A preferredmodification includes 2′-methoxyethoxy(2′-O—CH.sub.2CH.sub.2OCH.sub.3,also known as 2′-O-(2-methoxyethyl) or 2′-MOE) (Martin et al., Helv.Chim. Acta, 1995, 78, 486-504) i.e., an alkoxy-alkoxy group. A furtherpreferred modification includes 2′-dimethylaminooxyethoxy, i.e., aO(CH.sub.2).sub.2ON(CH.sub.3).sub.2 group, also known as 2′-DMAOE, asdescribed in examples hereinbelow, and 2′-dimethylaminoethoxyethoxy(also known in the art as 2′-O-dimethylaminoethoxyethyl or 2′-DMAEOE),i.e., 2′-O—CH.sub.2-O—CH.sub.2-N(CH.sub.2).sub.2, also described inexamples hereinbelow.

Other preferred modifications include 2′-methoxy(2′-OCH.sub.3),2′-aminopropoxy(2′-OCH.sub.2CH.sub.2CH.sub.2NH.sub.2) and 2′-fluoro(2′-F). Similar modifications may also be made at other positions on thedsRNA, particularly the 3′ position of the sugar on the 3′ terminalnucleotide or in 2′-5′ linked dsRNAs and the 5′ position of 5′ terminalnucleotide. DsRNAs may also have sugar mimetics such as cyclobutylmoieties in place of the pentofuranosyl sugar. Representative U.S.patents that teach the preparation of such modified sugar structuresinclude, but are not limited to, U.S. Pat. Nos. 4,981,957; 5,118,800;5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785;5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300;5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; and 5,700,920,certain of which are commonly owned with the instant application, andeach of which is herein incorporated by reference in its entirety.

dsRNAs may also include nucleobase (often referred to in the art simplyas “base”) modifications or substitutions. As used herein, “unmodified”or “natural” nucleobases include the purine bases adenine (A) andguanine (G), and the pyrimidine bases thymine (T), cytosine (C) anduracil (U). Modified nucleobases include other synthetic and naturalnucleobases such as 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine,xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkylderivatives of adenine and guanine, 2-propyl and other alkyl derivativesof adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine,5-halouracil and cytosine, 5-propynyl uracil and cytosine, 6-azo uracil,cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo,8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl anal other 8-substitutedadenines and guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyland other 5-substituted uracils and cytosines, 7-methylguanine and7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and7-daazaadenine and 3-deazaguanine and 3-deazaadenine. Furthernucleobases include those disclosed in U.S. Pat. No. 3,687,808, thosedisclosed in The Concise Encyclopedia Of Polymer Science AndEngineering, pages 858-859, Kroschwitz, J. L, ed. John Wiley & Sons,1990, these disclosed by Englisch et al., Angewandte Chemie,International Edition, 1991, 30, 613, and those disclosed by Sanghvi, YS., Chapter 15, DsRNA Research and Applications, pages 289-302, Crooke,S. T. and Lebleu, B., Ed., CRC Press, 1993. Certain of these nucleobasesare particularly useful for increasing the binding affinity of theoligomeric compounds of the invention. These include 5-substitutedpyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines,including 2-aminopropyladenine, 5-propynyluracil and 5-propynylcytosine.5-methylcytosine substitutions have been shown to increase nucleic acidduplex stability by 0.6-1.2.degree. C. (Sanghvi, Y. S., Crooke, S. T.and Lebleu, B., Eds., DsRNA Research and Applications, CRC Press, BocaRaton, 1993, pp. 276-278) and are presently preferred basesubstitutions, even more particularly when combined with2′-O-methoxyethyl sugar modifications.

Other nucleotide substitutions, such as “Universal” bases can beincorporated into siRNA duplexes to increase the number of targetsequences (or in this case, number of different Ebola strains) anyparticular siRNA might have complementarity to and activity against.Universal bases are non-canonical synthetic molecules that mimicstructures of traditional nucleotides (the genetic building blocks ofDNA and RNA). However, instead of selectively pairing according toWatson/Crick rules (A with T or U, C with G), universal bases ‘stack’equally well with all natural bases. Incorporating universal bases intosiRNAs may enable the siRNA to tolerate a mutation at that specific sitein its target mRNA. Thus, by decreasing the need for absolutecomplementarity between siRNA and its mRNA target, universal-basecontaining siRNAs may be an approach to (1) prevent drug resistancecaused by site-specific viral mutations and (2) create siRNAs able to bebroadly reactive across viral species with similar, but not absolutelyconserved, targets. Among the modifications that can be used asuniversal basaes are: 3-Nitropyrrole, 5-Nitroindole,Imidazole-4-Carboxamide, 2,4-difluorotoluyl, and Inosine.

Representative U.S. patents that teach the preparation of certain of theabove noted modified nucleobases as well as other modified nucleobasesinclude, but are not limited to, the above noted U.S. Pat. No.3,687,808, as well as U.S. Pat. Nos. 4,845,205; 5,130,30; 5,134,066;5,175,273; 5,367,066; 5,432,272; 5,457,187; 5,459,255; 5,484,908;5,502,177; 5,525,711; 5,552,540; 5,587,469; 5,594,121, 5,596,091;5,614,617; and 5,681,941, each of which is herein incorporated byreference, and U.S. Pat. No. 5,750,692, also herein incorporated byreference.

Another modification of the dsRNAs of the invention involves chemicallylinking to the dsRNA one or more moieties or conjugates which enhancethe activity, cellular distribution or cellular uptake of the dsRNA.Such moieties include but are not limited to lipid moieties such as acholesterol moiety (Letsinger et al., Proc. Natl. Acid. Sci. USA, 199,86, 6553-6556), cholic acid (Manoharan et al., Biorg. Med. Chem. Let.,1994 4 1053-1060), a thioether, e.g., beryl-S-tritylthiol (Manoharan etal., Ann. N.Y. Acad. Sci., 1992, 660, 306-309; Manoharan et al., Biorg.Med. Chem. Let., 1993, 3, 2765-2770), a thiocholesterol (Oberhauser etal., Nucl. Acids Res., 1992, 20, 533-538), an aliphatic chain, e.g.,dodecandiol or undecyl residues (Saison-Behmoaras et al., EMBO J, 1991,10, 1111-1118; Kabanov et al., FEBS Lett., 1990, 259, 327-330;Svinarchuk et al., Biochimie, 1993, 75, 49-54), a phospholipid, e.g.,di-hexadecyl-rac-glycerol or triethyl-ammonium1,2-di-O-hexadecyl-rac-glycero-3-Hphosphonate (Manoharan et al.,Tetrahedron Lett., 1995, 36, 3651-3654; Shea et al., Nucl. Acids Res.,1990, 18, 3777-3783), a polyamine or a polyethylene glycol chain(Manoharan et al., Nucleosides & Nucleotides, 1995, 14, 969-973), oradamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995, 36,3651-3654), a palmityl moiety (Mishra et al., Biochim. Biophys. Acta,1995, 1264, 229-237), or an octadecylamine orhexylamino-carbonyloxycholesterol moiety (Crooke et al., J. Pharmacol.Exp. Ther., 1996, 277, 923-937). Preferred conjugates will assist intargeting cells infected by Ebola virus such as dendritic cells andmacrophages which are involved in early stages of infection andepatocytes and other parenchymal cells which are involved in laterphases of the infection. Such conjugates include, but are not limitedto, mannose and folate conjugates.

Representative U.S. patents that teach the preparation of such dsRNAconjugates include, but are not limited to, U.S. Pat. Nos. 4,828,979;4,948,882; 5,218,105; 5,525,465; 5,541,313; 5,545,730; 5,552,538;5,578,717, 5,580,731; 5,591,584; 5,109,124; 5,118,802; 5,138,045;5,414,077; 5,486,603; 5,512,439; 5,578,718; 5,608,046; 4,587,044;4,605,735; 4,667,025; 4,762,779; 4,789,737; 4,824,941; 4,835,263;4,876,335; 4,904,582; 4,958,013; 5,082,830; 5,112,963; 5,214,136;5,082,830; 5,112,963; 5,214,136; 5,245,022; 5,254,469; 5,258,506;5,262,536; 5,272,250; 5,292,873; 5,317,098; 5,371,241, 5,391,723;5,416,203, 5,451,463; 5,510,475; 5,512,667; 5,514,785; 5,565,552;5,567,810; 5,574,142; 5,585,481; 5,587,371; 5,595,726; 5,597,696;5,599,923; 5,599,928 and 5,688,941, each of which is herein incorporatedby reference.

It is not necessary for all positions in a given compound to beuniformly modified, and in fact more than one of the aforementionedmodifications may be incorporated in a single compound or even at asingle nucleoside within an dsRNA. The present invention also includesdsRNA compounds which are chimeric compounds. “Chimeric” dsRNA compoundsor “chimeras,” in the context of this invention, are dsRNA compounds,particularly dsRNAs, which contain two or more chemically distinctregions, each made up of at least one monomer unit, i.e., a nucleotidein the case of an dsRNA compound. These dsRNAs typically contain atleast one region wherein the dsRNA is modified so as to confer upon thedsRNA increased resistance to nuclease degradation, increased cellularuptake, and/or increased binding affinity for the target nucleic acid.An additional region of the dsRNA may serve as a substrate for enzymescapable of cleaving RNA:DNA or RNA:RNA hybrids. By way of example, RNaseH is a cellular endonuclease which cleaves the RNA strand of an RNA:DNAduplex. Activation of RNase H, therefore, results in cleavage of the RNAtarget, thereby greatly enhancing the efficiency of dsRNA inhibition ofgene expression. Consequently, comparable results can often be obtainedwith shorter dsRNAs when chimeric dsRNAs are used, compared tophosphorothioate deoxydsRNAs hybridizing to the same target region.Cleavage of the RNA target can be routinely detected by gelelectrophoresis and, if necessary, associated nucleic acid hybridizationtechniques known in the art.

In certain instances, the dsRNA may be modified by a non-ligand group. Anumber of non-ligand molecules have been conjugated to dsRNAs in orderto enhance the activity, cellular distribution or cellular uptake of thedsRNA, and procedures for performing such conjugations are available inthe scientific literature. Such non-ligand moieties have included lipidmoieties, such as cholesterol (Letsinger et al., Proc. Natl. Acad. Sci.USA, 1989, 86:6553), cholic acid (Manoharan et al., Bioorg. Med. Chem.Lett., 1994, 4:1053), a thioether, e.g., hexyl-S-tritylthiol (Manoharanet al., Ann. N.Y. Acad. Sci., 1992, 660:306; Manoharan et al., Bioorg.Med. Chem. Let., 1993, 3:2765), a thiocholesterol (Oberhauser et al.,Nucl. Acids Res., 1992, 20:533), an aliphatic chain, e.g., dodecandiolor undecyl residues (Saison-Behmoaras et al., EMBO J., 1991, 10:111;Kabanov et al., FEBS Lett., 1990, 259:327; Svinarchuk et al., Biochimie,1993, 75:49), a phospholipid, e.g., di-hexadecyl-rac-glycerol ortriethylammonium 1,2-di-O-hexadecyl-rac-glycero-3-H-phosphonate(Manoharan et al., Tetrahedron Lett., 1995, 36:3651; Shea et al., Nucl.Acids Res., 1990, 18:3777), a polyamine or a polyethylene glycol chain(Manoharan et al., Nucleosides & Nucleotides, 1995, 14:969), oradamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995,36:3651), a palmityl moiety (Mishra et al., Biochim. Biophys. Acta,1995, 1264:229), or an octadecylamine orhexylamino-carbonyl-oxycholesterol moiety (Crooke et al., J. Pharmacol.Exp. Ther., 1996, 277:923). Representative United States patents thatteach the preparation of such dsRNA conjugates have been listed above.Typical conjugation protocols involve the synthesis of dsRNAs bearing anaminolinker at one or more positions of the sequence. The amino group isthen reacted with the molecule being conjugated using appropriatecoupling or activating reagents. The conjugation reaction may beperformed either with the dsRNA still bound to the solid support orfollowing cleavage of the dsRNA in solution phase. Purification of thedsRNA conjugate by HPLC typically affords the pure conjugate.

Vector Encoded RNAi Agents

The dsRNA of the invention can also be expressed from recombinant viralvectors intracellularly in vivo. The recombinant viral vectors of theinvention comprise sequences encoding the dsRNA of the invention and anysuitable promoter for expressing the dsRNA sequences. Suitable promotersinclude, for example, the U6 or H1 RNA pol III promoter sequences andthe cytomegalovirus promoter. Selection of other suitable promoters iswithin the skill in the art. The recombinant viral vectors of theinvention can also comprise inducible or regulatable promoters forexpression of the dsRNA in a particular tissue or in a particularintracellular environment. The use of recombinant viral vectors todeliver dsRNA of the invention to cells in vivo is discussed in moredetail below.

dsRNA of the invention can be expressed from a recombinant viral vectoreither as two separate, complementary RNA molecules, or as a single RNAmolecule with two complementary regions.

Any viral vector capable of accepting the coding sequences for the dsRNAmolecule(s) to be expressed can be used, for example vectors derivedfrom adenovirus (AV); adeno-associated virus (AAV); retroviruses (e.g,lentiviruses (LV), Rhabdoviruses, murine leukemia virus); herpes virus,and the like. The tropism of viral vectors can be modified bypseudotyping the vectors with envelope proteins or other surfaceantigens from other viruses, or by substituting different viral capsidproteins, as appropriate.

For example, lentiviral vectors of the invention can be pseudotyped withsurface proteins from vesicular stomatitis virus (VSV), rabies, Ebola,Mokola, and the like. AAV vectors of the invention can be made to targetdifferent cells by engineering the vectors to express different capsidprotein serotypes. For example, an AAV vector expressing a serotype 2capsid on a serotype 2 genome is called AAV 2/2. This serotype 2 capsidgene in the AAV 2/2 vector can be replaced by a serotype 5 capsid geneto produce an AAV 2/5 vector. Techniques for constructing AAV vectorswhich express different capsid protein serotypes are within the skill inthe art; see, e.g., Rabinowitz J E et al. (2002), J Virol 76:791-801,the entire disclosure of which is herein incorporated by reference.

Selection of recombinant viral vectors suitable for use in theinvention, methods for inserting nucleic acid sequences for expressingthe dsRNA into the vector, and methods of delivering the viral vector tothe cells of interest are within the skill in the art. See, for example,Dornburg R (1995), Gene Therap. 2: 301-310; Eglitis M A (1988),Biotechniques 6: 608-614; Miller A D (1990), Hum Gene Therap. 1: 5-14;Anderson W F (1998), Nature 392: 25-30; and Rubinson D A et al., Nat.Genet. 33: 401-406, the entire disclosures of which are hereinincorporated by reference.

Preferred viral vectors are those derived from AV and AAV. In aparticularly preferred embodiment, the dsRNA of the invention isexpressed as two separate, complementary single-stranded RNA moleculesfrom a recombinant AAV vector comprising, for example, either the U6 orH1 RNA promoters, or the cytomegalovirus (CMV) promoter.

A suitable AV vector for expressing the dsRNA of the invention, a methodfor constructing the recombinant AV vector, and a method for deliveringthe vector into target cells, are described in Xia H et al. (2002), Nat.Biotech. 20: 1006-1010.

Suitable AAV vectors for expressing the dsRNA of the invention, methodsfor constructing the recombinant AV vector, and methods for deliveringthe vectors into target cells are described in Samulski R et al. (1987),J. Virol. 61: 3096-3101; Fisher K J et al. (1996), J. Virol, 70:520-532; Samulski R et al. (1989), J. Virol. 63: 3822-3826; U.S. Pat.No. 5,252,479; U.S. Pat. No. 5,139,941; International Patent ApplicationNo. WO 94/13788; and International Patent Application No. WO 93/24641,the entire disclosures of which are herein incorporated by reference.

III. Pharmaceutical Compositions Comprising dsRNA

In one embodiment, the invention provides pharmaceutical compositionscomprising a dsRNA, as described herein, and a pharmaceuticallyacceptable carrier. The pharmaceutical composition comprising the dsRNAis useful for treating a disease or disorder associated with theexpression or activity of a gene from the Ebola virus and/or viralinfection, such as systemic hemorrhage and multi-organ failure. Suchpharmaceutical compositions are formulated based on the mode ofdelivery. One example is compositions that are formulated for systemicadministration via parenteral delivery.

The pharmaceutical compositions of the invention are administered indosages sufficient to inhibit expression of a gene from the Ebola virus.A maximum dosage of 5 mg dsRNA per kilogram body weight of recipient perday is sufficient to inhibit or completely suppress expression of a genefrom the Ebola virus.

In general, a suitable dose of dsRNA will be in the range of 0.01 to20.0 milligrams per kilogram body weight of the recipient per day, andoptimally in the range of 0.01 to 3 mg per kilogram body weight per day.The pharmaceutical composition may be administered once daily, or thedsRNA may be administered as two, three, or more sub-doses atappropriate intervals throughout the day or even using continuousinfusion or delivery through a controlled release formulation. In thatcase, the dsRNA contained in each sub-dose must be correspondinglysmaller in order to achieve the total daily dosage. The dosage unit canalso be compounded for delivery over several days, e.g., using aconventional sustained release formulation which provides sustainedrelease of the dsRNA over a several day period. In this embodiment, thedosage unit contains a corresponding multiple of the daily dose.

The skilled artisan will appreciate that certain factors may influencethe dosage and timing required to effectively treat a subject, includingbut not limited to the severity of the disease or disorder, previoustreatments, the general health and/or age of the subject, and otherdiseases present. Moreover, treatment of a subject with atherapeutically effective amount of a composition can include a singletreatment or a series of treatments. Estimates of effective dosages andin vivo half-lives for the individual dsRNAs encompassed by theinvention can be made using conventional methodologies or on the basisof in vivo testing using an appropriate animal model, as describedelsewhere herein.

Advances in animal modeling has generated a number of Ebola infectionmodels (mouse, guinea pig, and non-human primate) that reproduce all themajor pathologies associated with human Ebola infection (reviewed inWarfield, K. L. et al. (2006) “Chapter 13: Viral Hemorrhagic Fevers” inBiodefense:Research Methodology and Animal Models, pages 227-258, J. R.Swearengen, Ed., Taylor & Francis, Boca Raton). Such models are used forin vivo testing of dsRNA, as well as for determining a therapeuticallyeffective dose.

The present invention also includes pharmaceutical compositions andformulations which include the dsRNA compounds of the invention. Thepharmaceutical compositions of the present invention may be administeredin a number of ways depending upon whether local or systemic treatmentis desired and upon the area to be treated. Administration may betopical, pulmonary, e.g., by inhalation or insufflation of powders oraerosols, including by nebulizer; intratracheal, intranasal, epidermaland transdermal), oral or parenteral. Parenteral administration includesintravenous, intraarterial, subcutaneous, intraperitoneal orintramuscular injection or infusion.

Compositions and formulations for oral administration include powders orgranules, microparticulates, nanoparticulates, suspensions or solutionsin water or non-aqueous media, capsules, gel capsules, sachets, tabletsor minitablets. Thickeners, flavoring agents, diluents, emulsifiers,dispersing aids or binders may be desirable.

Compositions and formulations for parenteral, intrathecal orintraventricular administration may include sterile aqueous solutionswhich may also contain buffers, diluents and other suitable additivessuch as, but not limited to, penetration enhancers, carrier compoundsand other pharmaceutically acceptable carriers or excipients.

Pharmaceutical compositions of the present invention include, but arenot limited to, solutions, emulsions, and liposome-containingformulations. These compositions may be generated from a variety ofcomponents that include, but are not limited to, preformed liquids,self-emulsifying solids and self-emulsifying semisolids.

The pharmaceutical formulations of the present invention, which mayconveniently be presented in unit dosage form, may be prepared accordingto conventional techniques well known in the pharmaceutical industry.Such techniques include the step of bringing into association the activeingredients with the pharmaceutical carrier(s) or excipient(s). Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association the active ingredients with liquid carriers orfinely divided solid carriers or both, and then, if necessary, shapingthe product.

Liposomes

There are many organized surfactant structures besides microemulsionsthat have been studied and used for the formulation of drugs. Theseinclude monolayers, micelles, bilayers and vesicles. Vesicles, such asliposomes, have attracted great interest because of their specificityand the duration of action they offer from the standpoint of drugdelivery. Liposomal delivery systems have been used to effectivelydeliver siRNA in vivo and silence genes in hepatocytes [Zimmermann etal. (2006) Nature, 441:111-114]. Such siRNA-liposomal formulations havealso been used for therapeutic benefit in animal models of dyslipidemias[Zimmermann et al. (2006) Nature, 441:111-114], HBV infection [Morrisseyet al. (2005) Nature Biotech 23:1002-1007], Ebola infection [Geisbert,et al. (2006) The Journal of Infectious Diseases, 193:1650-1657], andrheumatoid arthritis [Khoury et al. (2006) Arthritis & Rheumatism,54:1867-1877]. As used in the present invention, the term “liposome”means a vesicle composed of amphiphilic lipids arranged in a sphericalbilayer or bilayers.

Liposomes are unilamellar or multilamellar vesicles which have amembrane formed from a lipophilic material and an aqueous interior. Theaqueous portion contains the composition to be delivered. Cationicliposomes possess the advantage of being able to fuse to the cell wall.Non-cationic liposomes, although not able to fuse as efficiently withthe cell wall, are phagocytosed by macrophages and other cells in vivo.

Further advantages of liposomes include; liposomes obtained from naturalphospholipids are biocompatible and biodegradable; liposomes canincorporate a wide range of water and lipid soluble drugs; liposomes canprotect encapsulated drugs in their internal compartments frommetabolism and degradation (Rosoff, in Pharmaceutical Dosage Forms,Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., NewYork, N.Y., volume 1, p. 245). Important considerations in thepreparation of liposome formulations are the lipid surface charge,vesicle size and the aqueous volume of the liposomes.

Liposomes are useful for the transfer and delivery of active ingredientsto the site of action. Because the liposomal membrane is structurallysimilar to biological membranes, when liposomes are applied to a tissue,the liposomes start to merge with the cellular membranes and as themerging of the liposome and cell progresses, the liposomal contents areemptied into the cell where the active agent may act.

Liposomal formulations have been the focus of extensive investigation asthe mode of delivery for many drugs. There is growing evidence that fortopical administration, liposomes present several advantages over otherformulations. Such advantages include reduced side-effects related tohigh systemic absorption of the administered drug, increasedaccumulation of the administered drug at the desired target, and theability to administer a wide variety of drugs, both hydrophilic andhydrophobic, into the skin.

Several reports have detailed the ability of liposomes to deliver agentsincluding high-molecular weight DNA into the skin. Compounds includinganalgesics, antibodies, hormones and high-molecular weight DNAs havebeen administered to the skin. The majority of applications resulted inthe targeting of the upper epidermis

Liposomes fall into two broad classes. Cationic liposomes are positivelycharged liposomes which interact with the negatively charged DNAmolecules to form a stable complex. The positively charged DNA/liposomecomplex binds to the negatively charged cell surface and is internalizedin an endosome. Due to the acidic pH within the endosome, the liposomesare ruptured, releasing their contents into the cell cytoplasm (Wang etal., Biochem. Biophys. Res. Commun., 1987, 147, 980-985).

Liposomes which are pH-sensitive or negatively-charged, entrap DNArather than complex with it. Since both the DNA and the lipid aresimilarly charged, repulsion rather than complex formation occurs.Nevertheless, some DNA is entrapped within the aqueous interior of theseliposomes. pH-sensitive liposomes have been used to deliver DNA encodingthe thymidine kinase gene to cell monolayers in culture. Expression ofthe exogenous gene was detected in the target cells (Zhou et al.,Journal of Controlled Release, 1992, 19, 269-274).

One major type of liposomal composition includes phospholipids otherthan naturally-derived phosphatidylcholine. Neutral liposomecompositions, for example, can be formed from dimyristoylphosphatidylcholine (DMPC) or dipalmitoyl phosphatidylcholine (DPPC).Anionic liposome compositions generally are formed from dimyristoylphosphatidylglycerol, while anionic fusogenic liposomes are formedprimarily from dioleoyl phosphatidylethanolamine (DOPE). Another type ofliposomal composition is formed from phosphatidylcholine (PC) such as,for example, soybean PC, and egg PC. Another type is formed frommixtures of phospholipid and/or phosphatidylcholine and/or cholesterol.

Liposomes also include “sterically stabilized” liposomes, a term which,as used herein, refers to liposomes comprising one or more specializedlipids that, when incorporated into liposomes, result in enhancedcirculation lifetimes relative to liposomes lacking such specializedlipids. Examples of sterically stabilized liposomes are those in whichpart of the vesicle-forming lipid portion of the liposome (A) comprisesone or more glycolipids, such as monosialoganglioside G.sub.M1, or (B)is derivatized with one or more hydrophilic polymers, such as apolyethylene glycol (PEG) moiety. While not wishing to be bound by anyparticular theory, it is thought in the art that, at least forsterically stabilized liposomes containing gangliosides, sphingomyelin,or PEG-derivatized lipids, the enhanced circulation half-life of thesesterically stabilized liposomes derives from a reduced uptake into cellsof the reticuloendothelial system (RES) (Allen et al., FEBS Letters,1987, 223, 42; Wu et al., Cancer Research, 1993, 53, 3765). Variousliposomes comprising one or more glycolipids are known in the art.Papahadjopoulos et al. (Ann. N.Y. Acad. Sci., 1987, 507, 64) reportedthe ability of monosialoganglioside G.sub.M1, galactocerebroside sulfateand phosphatidylinositol to improve blood half-lives of liposomes. Thesefindings were expounded upon by Gabizon et al. (Proc. Natl. Acad. Sci.U.S.A., 1988, 85, 6949). U.S. Pat. No. 4,837,028 and WO 88/04924, bothto Allen et al., disclose liposomes comprising (1) sphingomyelin and (2)the ganglioside G.sub.M1 or a galactocerebroside sulfate ester. U.S.Pat. No. 5,543,152 (Webb et al.) discloses liposomes comprisingsphingomyelin. Liposomes comprising1,2-sn-dimyristoylphosphat-idylcholine are disclosed in WO 97/13499 (Limet al).

Many liposomes comprising lipids derivatized with one or morehydrophilic polymers, and methods of preparation thereof, are known inthe art. Sunamoto et al. (Bull. Chem. Soc. Jpn., 1980, 53, 2778)described liposomes comprising a nonionic detergent, 2C.sub.1215G, thatcontains a PEG moiety. Illum et al. (FEBS Lett., 1984, 167, 79) notedthat hydrophilic coating of polystyrene particles with polymeric glycolsresults in significantly enhanced blood half-lives. Syntheticphospholipids modified by the attachment of carboxylic groups ofpolyalkylene glycols (e.g., PEG) are described by Sears (U.S. Pat. Nos.4,426,330 and 4,534,899). Klibanov et al. (FEBS Lett., 1990, 268, 235)described experiments demonstrating that liposomes comprisingphosphatidylethanolamine (PE) derivatized with PEG or PEG stearate havesignificant increases in blood circulation half-lives. Blume et al.(Biochimica et Biophysica Acta, 1990, 1029, 91) extended suchobservations to other PEG-derivatized phospholipids, e.g., DSPE-PEG,formed from the combination of distearoylphosphatidylethanolamine (DSPE)and PEG. Liposomes having covalently bound PEG moieties on theirexternal surface are described in European Patent No. EP 0 445 131 B1and WO 90/04384 to Fisher. Liposome compositions containing 1-20 molepercent of PE derivatized with PEG, and methods of use thereof, aredescribed by Woodle et al. (U.S. Pat. Nos. 5,013,556 and 5,356,633) andMartin et al. (U.S. Pat. No. 5,213,804 and European Patent No. EP 0 496813 B1). Liposomes comprising a number of other lipid-polymer conjugatesare disclosed in WO 91/05545 and U.S. Pat. No. 5,225,212 (both to Martinet al.) and in WO 94/20073 (Zalipsky et al.) Liposomes comprisingPEG-modified ceramide lipids are described in WO 96/10391 (Choi et al).U.S. Pat. No. 5,540,935 (Miyazaki et al.) and U.S. Pat. No. 5,556,948(Tagawa et al.) describe PEG-containing liposomes that can be furtherderivatized with functional moieties on their surfaces.

Liposomes and other nanoparticles have also been designed which containspecific targeting molecules. Targeting molecules used for siRNAdelivery in vivo have included integrin-binding RGD peptides[Schiffelers et al. (2004) Nucleic Acids Research 32:e149), anisamide[Li and Huang (2006) Molecular Pharmaceutics 3:579-588] and folate[Hu-Lieskovan et al. (2005) Cancer Research 65:8984-8992]. For deliveryto myeloid and dendritic cells which are presumed to be important inearly Ebola infection, incorporation of targeting agents such as mannoseand folate into liposomes and nanoparticles may improve both siRNAdelivery and therapeutic effect. Mannose-conjugated oligonucleotideshave been shown to specifically improve delivery to myeloid cells[Rojanasakul et al. (1997) Journal of Biological Chemistry272:3910-3914; Diebold et al (2002) Somatic Cell and Molecular Genetics27:65-73] and mannosylated liposomes are effective targeting agents invivo [Diebold et al. (2002) Somatic Cell and Molecular Genetics27:65-73; Hattori et al. (2006) Journal Gene Medicine 8:824-834; Hattoriet al. (2006) Journal of Pharmacology and Experimental Therapeutics318:828-834]. Folate conjugation has proven an effective deliveryvehicle in a wide variety of contexts [reviewed in Hilgenbrink and Low(2005) Journal Pharmaceutical Sciences 94:2135-2146] and incorporationof folate into liposomes is possible using commercially availablereagents such as DSPE-PEG(2000) Folate (Avanti Polar Lipids, Alabaster,Ala.). A limited number of liposomes comprising nucleic acids are knownin the art. WO 96/40062 to Thierry et al. discloses methods forencapsulating high molecular weight nucleic acids in liposomes. U.S.Pat. No. 5,264,221 to Tagawa et al. discloses protein-bonded liposomesand asserts that the contents of such liposomes may include an dsRNARNA. U.S. Pat. No. 5,665,710 to Rahman et al. describes certain methodsof encapsulating oligodeoxynucleotides in liposomes. WO 97/04787 to Loveet al. discloses liposomes comprising dsRNA targeted to the raf gene.

Transfersomes are yet another type of liposomes, and are highlydeformable lipid aggregates which are attractive candidates for drugdelivery vehicles. Transfersomes may be described as lipid dropletswhich are so highly deformable that they are easily able to penetratethrough pores which are smaller than the droplet. Transfersomes areadaptable to the environment in which they are used, e.g. they areself-optimizing (adaptive to the shape of pores in the skin),self-repairing, frequently reach their targets without fragmenting, andoften self-loading. To make transfersomes it is possible to add surfaceedge-activators, usually surfactants, to a standard liposomalcomposition. Transfersomes have been used to deliver serum albumin tothe skin. The transfersome-mediated delivery of serum albumin has beenshown to be as effective as subcutaneous injection of a solutioncontaining serum albumin.

Surfactants find wide application in formulations such as emulsions(including microemulsions) and liposomes. The most common way ofclassifying and ranking the properties of the many different types ofsurfactants, both natural and synthetic, is by the use of thehydrophile/lipophile balance (HLB). The nature of the hydrophilic group(also known as the “head”) provides the most useful means forcategorizing the different surfactants used in formulations (Rieger, inPharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988,p. 285).

If the surfactant molecule is not ionized, it is classified as anonionic surfactant. Nonionic surfactants find wide application inpharmaceutical and cosmetic products and are usable over a wide range ofpH values. In general their HLB values range from 2 to about 18depending on their structure. Nonionic surfactants include nonionicesters such as ethylene glycol esters, propylene glycol esters, glycerylesters, polyglyceryl esters, sorbitan esters, sucrose esters, andethoxylated esters. Nonionic alkanolamides and ethers such as fattyalcohol ethoxylates, propoxylated alcohols, and ethoxylated/propoxylatedblock polymers are also included in this class. The polyoxyethylenesurfactants are the most popular members of the nonionic surfactantclass.

If the surfactant molecule carries a negative charge when it isdissolved or dispersed in water, the surfactant is classified asanionic. Anionic surfactants include carboxylates such as soaps, acyllactylates, acyl amides of amino acids, esters of sulfuric acid such asalkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkylbenzene sulfonates, acyl isethionates, acyl taurates andsulfosuccinates, and phosphates. The most important members of theanionic surfactant class are the alkyl sulfates and the soaps.

If the surfactant molecule carries a positive charge when it isdissolved or dispersed in water, the surfactant is classified ascationic. Cationic surfactants include quaternary ammonium salts andethoxylated amines. The quaternary ammonium salts are the most usedmembers of this class.

If the surfactant molecule has the ability to carry either a positive ornegative charge, the surfactant is classified as amphoteric. Amphotericsurfactants include acrylic acid derivatives, substituted alkylamides,N-alkylbetaines and phosphatides.

The use of surfactants in drug products, formulations and in emulsionshas been reviewed (Rieger, in Pharmaceutical Dosage Forms, MarcelDekker, Inc., New York, N.Y., 1988, p. 285).

Agents that enhance uptake of dsRNAs at the cellular level may also beadded to the pharmaceutical and other compositions of the presentinvention. For example, cationic lipids, such as lipofectin (Junichi etal, U.S. Pat. No. 5,705,188), cationic glycerol derivatives, andpolycationic molecules, such as polylysine (Lollo et al., PCTApplication WO 97/30731), are also known to enhance the cellular uptakeof dsRNAs.

Other agents may be utilized to enhance the penetration of theadministered nucleic acids, including glycols such as ethylene glycoland propylene glycol, pyrrols such as 2-pyrrol, azones, and terpenessuch as limonene and menthone.

Carriers

Certain compositions of the present invention also incorporate carriercompounds in the formulation. As used herein, “carrier compound” or“carrier” can refer to a nucleic acid, or analog thereof, which is inert(i.e., does not possess biological activity per se) but is recognized asa nucleic acid by in vivo processes that reduce the bioavailability of anucleic acid having biological activity by, for example, degrading thebiologically active nucleic acid or promoting its removal fromcirculation. The coadministration of a nucleic acid and a carriercompound, typically with an excess of the latter substance, can resultin a substantial reduction of the amount of nucleic acid recovered inthe liver, kidney or other extracirculatory reservoirs, presumably dueto competition between the carrier compound and the nucleic acid for acommon receptor. For example, the recovery of a partiallyphosphorothioate dsRNA in hepatic tissue can be reduced when it iscoadministered with polyinosinic acid, dextran sulfate, polycytidic acidor 4-acetamido-4′ isothiocyano-stilbene-2,2′-disulfonic acid (Miyao etal., DsRNA Res. Dev., 1995, 5, 115-121; Takakura et al., DsRNA & Nucl.Acid Drug Dev., 1996, 6, 177-183.

Excipients

In contrast to a carrier compound, a “pharmaceutical carrier” or“excipient” is a pharmaceutically acceptable solvent, suspending agentor any other pharmacologically inert vehicle for delivering one or morenucleic acids to an animal. The excipient may be liquid or solid and isselected, with the planned manner of administration in mind, so as toprovide for the desired bulk, consistency, etc., when combined with anucleic acid and the other components of a given pharmaceuticalcomposition. Typical pharmaceutical carriers include, but are notlimited to, binding agents (e.g., pregelatinized maize starch,polyvinylpyrrolidone or hydroxypropyl methylcellulose, etc.); fillers(e.g., lactose and other sugars, microcrystalline cellulose, pectin,gelatin, calcium sulfate, ethyl cellulose, polyacrylates or calciumhydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc,silica, colloidal silicon dioxide, stearic acid, metallic stearates,hydrogenated vegetable oils, corn starch, polyethylene glycols, sodiumbenzoate, sodium acetate, etc.); disintegrants (e.g., starch, sodiumstarch glycolate, etc.); and wetting agents (e.g., sodium laurylsulphate, etc).

Pharmaceutically acceptable organic or inorganic excipient suitable fornon-parenteral administration which do not deleteriously react withnucleic acids can also be used to formulate the compositions of thepresent invention. Suitable pharmaceutically acceptable carriersinclude, but are not limited to, water, salt solutions, alcohols,polyethylene glycols, gelatin, lactose, amylose, magnesium stearate,talc, silicic acid, viscous paraffin, hydroxymethylcellulose,polyvinylpyrrolidone and the like.

Suitable pharmaceutically acceptable excipients include, but are notlimited to, water, salt solutions, alcohol, polyethylene glycols,gelatin, lactose, amylose, magnesium stearate, talc, silicic acid,viscous paraffin, hydroxymethylcellulose, polyvinylpyrrolidone and thelike.

Other Components

The compositions of the present invention may additionally contain otheradjunct components conventionally found in pharmaceutical compositions,at their art-established usage levels. Thus, for example, thecompositions may contain additional, compatible, pharmaceutically-activematerials such as, for example, antipruritics, astringents, localanesthetics or anti-inflammatory agents, or may contain additionalmaterials useful in physically formulating various dosage forms of thecompositions of the present invention, such as dyes, flavoring agents,preservatives, antioxidants, opacifiers, thickening agents andstabilizers. However, such materials, when added, should not undulyinterfere with the biological activities of the components of thecompositions of the present invention. The formulations can besterilized and, if desired, mixed with auxiliary agents, e.g.,lubricants, preservatives, stabilizers, wetting agents, emulsifiers,salts for influencing osmotic pressure, buffers, colorings, flavoringsand/or aromatic substances and the like which do not deleteriouslyinteract with the nucleic acid(s) of the formulation.

Aqueous suspensions may contain substances which increase the viscosityof the suspension including, for example, sodium carboxymethylcellulose,sorbitol and/or dextran. The suspension may also contain stabilizers.

Certain embodiments of the invention provide pharmaceutical compositionscontaining (a) one or more antisense compounds and (b) one or more otherchemotherapeutic agents which function by a non-antisense mechanism.Examples of such chemotherapeutic agents include but are not limited todaunorubicin, daunomycin, dactinomycin, doxorubicin, epirubicin,idarubicin, esorubicin, bleomycin, mafosfamide, ifosfamide, cytosinearabinoside, bis-chloroethylnitrosurea, busulfan, mitomycin C,actinomycin D, mithramycin, prednisone, hydroxyprogesterone,testosterone, tamoxifen, dacarbazine, procarbazine, hexamethylmelamine,pentamethylmelamine, mitoxantrone, amsacrine, chlorambucil,methylcyclohexylnitrosurea, nitrogen mustards, melphalan,cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine,5-azacytidine, hydroxyurea, deoxycoformycin,4-hydroxyperoxycyclophosphor-amide, 5-fluorouracil (5-FU),5-fluorodeoxyuridine (5-FUdR), methotrexate (MTX), colchicine, taxol,vincristine, vinblastine, etoposide (VP-16), trimetrexate, irinotecan,topotecan, gemcitabine, teniposide, cisplatin and diethylstilbestrol(DES). See, generally, The Merck Manual of Diagnosis and Therapy, 15thEd. 1987, pp. 1206-1228, Berkow et al., eds., Rahway, N.J. When usedwith the compounds of the invention, such chemotherapeutic agents may beused individually (e.g., 5-FU and oligonucleotide), sequentially (e.g.,5-FU and oligonucleotide for a period of time followed by MTX andoligonucleotide), or in combination with one or more other suchchemotherapeutic agents (e.g., 5-FU, MTX and oligonucleotide, or 5-FU,radiotherapy and oligonucleotide). Anti-inflammatory drugs, includingbut not limited to nonsteroidal anti-inflammatory drugs andcorticosteroids, and antiviral drugs, including but not limited toribivirin, vidarabine, acyclovir and ganciclovir, may also be combinedin compositions of the invention. See, generally, The Merck Manual ofDiagnosis and Therapy, 15th Ed., Berkow et al., eds., 1987, Rahway,N.J., pages 2499-2506 and 46-49, respectively). Other non-antisensechemotherapeutic agents are also within the scope of this invention. Twoor more combined compounds may be used together or sequentially.

Toxicity and therapeutic efficacy of such compounds can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the LD50 (the dose lethal to 50% of thepopulation) and the ED50 (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and it can be expressed as the ratio LD50/ED50.Compounds which exhibit high therapeutic indices are preferred.

The data obtained from cell culture assays and animal studies can beused in formulation a range of dosage for use in humans. The dosage ofcompositions of the invention lies generally within a range ofcirculating concentrations that include the ED50 with little or notoxicity. The dosage may vary within this range depending upon thedosage form employed and the route of administration utilized. For anycompound used in the method of the invention, the therapeuticallyeffective dose can be estimated initially from cell culture assays. Adose may be formulated in animal models to achieve a circulating plasmaconcentration range of the compound or, when appropriate, of thepolypeptide product of a target sequence (e.g., achieving a decreasedconcentration of the polypeptide) that includes the IC50 (i.e., theconcentration of the test compound which achieves a half-maximalinhibition of symptoms) as determined in cell culture. Such informationcan be used to more accurately determine useful doses in humans. Levelsin plasma may be measured, for example, by high performance liquidchromatography.

In addition to their administration individually or as a plurality, asdiscussed above, the dsRNAs of the invention can be administered incombination with other known agents effective in treatment ofpathological processes mediated by Ebola expression. In any event, theadministering physician can adjust the amount and timing of dsRNAadministration on the basis of results observed using standard measuresof efficacy known in the art or described herein.

Methods for Treating Diseases Caused by Infection with the Ebola Virus

The invention relates in particular to the use of a dsRNA or apharmaceutical composition prepared therefrom for the treatment orprevention of pathological conditions associated with Ebola infection,e.g., systemic hemorrhage and multi-organ failure.

The invention furthermore relates to the use of an dsRNA or apharmaceutical composition thereof for treating systemic hemorrhage andmulti-organ failure in combination with other pharmaceuticals and/orother therapeutic methods, e.g., with known pharmaceuticals and/or knowntherapeutic methods, such as, for example, those which are currentlyemployed for treating viral infection and systemic hemorrhage.Preference is given to a combination with interferon or other antiviralagents.

Methods for Inhibiting Expression of a Gene from the Ebola Virus

In yet another aspect, the invention provides a method for inhibitingthe expression of a gene from the Ebola virus in a mammal. The methodcomprises administering a composition of the invention to the mammalsuch that expression of the target Ebola genome is silenced. Because oftheir high specificity, the dsRNAs of the invention specifically targetRNAs (primary or processed) of the target Ebola gene. Compositions andmethods for inhibiting the expression of these Ebola genes using dsRNAscan be performed as described elsewhere herein.

In one embodiment, the method comprises administering a compositioncomprising a dsRNA, wherein the dsRNA comprises a nucleotide sequencewhich is complementary to at least a part of an RNA transcript of a genefrom the Ebola virus, to the mammal to be treated. When the organism tobe treated is a mammal such as a human, the composition may beadministered by any means known in the art including, but not limited tooral or parenteral routes, including intravenous, intraperitoneal,intramuscular, subcutaneous, transdermal, airway (aerosol), nasal,administration. In some embodiments, the compositions are administeredby intravenous infusion or injection.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the invention, suitable methods and materials aredescribed below. All publications, patent applications, patents, andother references mentioned herein are incorporated by reference in theirentirety. In case of conflict, the present specification, includingdefinitions, will control. In addition, the materials, methods, andexamples are illustrative only and not intended to be limiting.

EXAMPLES Example 1 dsRNA Synthesis

Source of Reagents

Where the source of a reagent is not specifically given herein, suchreagent may be obtained from any supplier of reagents for molecularbiology at a quality/purity standard for application in molecularbiology.

siRNA Synthesis

Single-stranded RNAs were produced by solid phase synthesis on a scaleof 1 μmole using an Expedite 8909 synthesizer (Applied Biosystems,Applera Deutschland GmbH, Darmstadt, Germany) and controlled pore glass(CPG, 500Å, Proligo Biochemie GmbH, Hamburg, Germany) as solid support.RNA and RNA containing 2′-O-methyl nucleotides were generated by solidphase synthesis employing the corresponding phosphoramidites and2′-O-methyl phosphoramidites, respectively (Proligo Biochemie GmbH,Hamburg, Germany). These building blocks were incorporated at selectedsites within the sequence of the oligoribonucleotide chain usingstandard nucleoside phosphoramidite chemistry such as described inCurrent protocols in nucleic acid chemistry, Beaucage, S. L. et al.(Edrs.), John Wiley & Sons, Inc., New York, N.Y., USA. Phosphorothioatelinkages were introduced by replacement of the iodine oxidizer solutionwith a solution of the Beaucage reagent (Chruachem Ltd, Glasgow, UK) inacetonitrile (1%). Further ancillary reagents were obtained fromMallinckrodt Baker (Griesheim, Germany).

Deprotection and purification of the crude oligoribonucleotides by anionexchange HPLC were carried out according to established procedures.Yields and concentrations were determined by UV absorption of a solutionof the respective RNA at a wavelength of 260 nm using a spectralphotometer (DU 640B, Beckman Coulter GmbH, Unterschleiβheim, Germany).Double stranded RNA was generated by mixing an equimolar solution ofcomplementary strands in annealing buffer (20 mM sodium phosphate, pH6.8; 100 mM sodium chloride), heated in a water bath at 85-90° C. for 3minutes and cooled to room temperature over a period of 3-40 hours. Theannealed RNA solution was stored at −20° C. until use.

For the synthesis of 3′-cholesterol-conjugated siRNAs (herein referredto as -Chol-3′), an appropriately modified solid support was used forRNA synthesis. The modified solid support was prepared as follows:

Diethyl-2-azabutane-1,4-dicarboxylate AA

A 4.7 M aqueous solution of sodium hydroxide (50 mL) was added into astirred, ice-cooled solution of ethyl glycinate hydrochloride (32.19 g,0.23 mole) in water (50 mL). Then, ethyl acrylate (23.1 g, 0.23 mole)was added and the mixture was stirred at room temperature untilcompletion of the reaction was ascertained by TLC. After 19 h thesolution was partitioned with dichloromethane (3×100 mL). The organiclayer was dried with anhydrous sodium sulfate, filtered and evaporated.The residue was distilled to afford AA (28.8 g, 61%).

3-{Ethoxycarbonylmethyl-[6-(9H-fluoren-9-ylmethoxycarbonyl-amino)-hexanoyl]-amino}-propionicacid ethyl ester AB

Fmoc-6-amino-hexanoic acid (9.12 g, 25.83 mmol) was dissolved indichloromethane (50 mL) and cooled with ice. Diisopropylcarbodiimde(3.25 g, 3.99 mL, 25.83 mmol) was added to the solution at 0° C. It wasthen followed by the addition of Diethyl-azabutane-1,4-dicarboxylate (5g, 24.6 mmol) and dimethylamino pyridine (0.305 g, 2.5 mmol). Thesolution was brought to room temperature and stirred further for 6 h.Completion of the reaction was ascertained by TLC. The reaction mixturewas concentrated under vacuum and ethyl acetate was added to precipitatediisopropyl urea. The suspension was filtered. The filtrate was washedwith 5% aqueous hydrochloric acid, 5% sodium bicarbonate and water. Thecombined organic layer was dried over sodium sulfate and concentrated togive the crude product which was purified by column chromatography (50%EtOAC/Hexanes) to yield 11.87 g (88%) of AB.

3-[(6-Amino-hexanoyl)-ethoxycarbonylmethyl-amino]-propionic acid ethylester AC

3-{Ethoxycarbonylmethyl-[6-(9H-fluoren-9-ylmethoxycarbonylamino)-hexanoyl]-amino}-propionicacid ethyl ester AB (11.5 g, 21.3 mmol) was dissolved in 20% piperidinein dimethylformamide at 0° C. The solution was continued stirring for 1h. The reaction mixture was concentrated under vacuum, water was addedto the residue, and the product was extracted with ethyl acetate. Thecrude product was purified by conversion into its hydrochloride salt.

3-({6-[17-(1,5-Dimethyl-hexyl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro1H-cyclopenta[a]phenanthren-3-yloxycarbonylamino]-hexanoyl}ethoxycarbonylmethyl-amino)-propionicacid ethyl ester AD

The hydrochloride salt of3-[(6-Amino-hexanoyl)-ethoxycarbonylmethyl-amino]-propionic acid ethylester AC (4.7 g, 14.8 mmol) was taken up in dichloromethane. Thesuspension was cooled to 0° C. on ice. To the suspensiondiisopropylethylamine (3.87 g, 5.2 mL, 30 mmol) was added. To theresulting solution cholesteryl chloroformate (6.675 g, 14.8 mmol) wasadded. The reaction mixture was stirred overnight. The reaction mixturewas diluted with dichloromethane and washed with 10% hydrochloric acid.The product was purified by flash chromatography (10.3 g, 92%).

1-{6-[17-(1,5-Dimethyl-hexyl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yloxycarbonylamino]-hexanoyl}-4-oxo-pyrrolidine-3-carboxylicacid ethyl ester AE

Potassium t-butoxide (1.1 g, 9.8 mmol) was slurried in 30 mL of drytoluene. The mixture was cooled to 0° C. on ice and 5 g (6.6 mmol) ofdiester AD was added slowly with stirring within 20 mins. Thetemperature was kept below 5° C. during the addition. The stirring wascontinued for 30 mins at 0° C. and 1 mL of glacial acetic acid wasadded, immediately followed by 4 g of NaH₂PO₄.H₂O in 40 mL of water Theresultant mixture was extracted twice with 100 mL of dichloromethaneeach and the combined organic extracts were washed twice with 10 mL ofphosphate buffer each, dried, and evaporated to dryness. The residue wasdissolved in 60 mL of toluene, cooled to 0° C. and extracted with three50 mL portions of cold pH 9.5 carbonate buffer. The aqueous extractswere adjusted to pH 3 with phosphoric acid, and extracted with five 40mL portions of chloroform which were combined, dried and evaporated todryness. The residue was purified by column chromatography using 25%ethylacetate/hexane to afford 1.9 g of b-ketoester (39%).

[6-(3-Hydroxy-4-hydroxymethyl-pyrrolidin-1-yl)-6-oxo-hexyl]-carbamicacid17-(1,5-dimethyl-hexyl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ylester AF

Methanol (2 mL) was added dropwise over a period of 1 h to a refluxingmixture of b-ketoester AE (1.5 g, 2.2 mmol) and sodium borohydride(0.226 g, 6 mmol) in tetrahydrofuran (10 mL). Stirring was continued atreflux temperature for 1 h. After cooling to room temperature, 1N HCl(12.5 mL) was added, the mixture was extracted with ethylacetate (3×40mL). The combined ethylacetate layer was dried over anhydrous sodiumsulfate and concentrated under vacuum to yield the product which waspurified by column chromatography (10% MeOH/CHCl₃) (89%).

(6-{3-[Bis-(4-methoxy-phenyl)-phenyl-methoxymethyl]-4-hydroxy-pyrrolidin-1-yl}-6-oxo-hexyl)-carbamicacid17-(1,5-dimethyl-hexyl)-10,13-dimethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-ylester AG

Diol AF (1.25 μm 1.994 mmol) was dried by evaporating with pyridine (2×5mL) in vacuo. Anhydrous pyridine (10 mL) and4,4′-dimethoxytritylchloride (0.724 g, 2.13 mmol) were added withstirring. The reaction was carried out at room temperature overnight.The reaction was quenched by the addition of methanol. The reactionmixture was concentrated under vacuum and to the residue dichloromethane(50 mL) was added. The organic layer was washed with 1M aqueous sodiumbicarbonate. The organic layer was dried over anhydrous sodium sulfate,filtered and concentrated. The residual pyridine was removed byevaporating with toluene. The crude product was purified by columnchromatography (2% MeOH/Chloroform, Rf=0.5 in 5% MeOH/CHCl₃) (1.75 g,95%).

Succinic acidmono-(4-[bis-(4-methoxy-phenyl)-phenyl-methoxymethyl]-1-{6-[17-(1,5-dimethyl-hexyl)-10,13-dimethyl2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1Hcyclopenta[a]phenanthren-3-yloxycarbonylamino]-hexanoyl}-pyrrolidin-3-yl)ester AH

Compound AG (1.0 g, 1.05 mmol) was mixed with succinic anhydride (0.150g, 1.5 mmol) and DMAP (0.073 g, 0.6 mmol) and dried in a vacuum at 40°C. overnight. The mixture was dissolved in anhydrous dichloroethane (3mL), triethylamine (0.318 g, 0.440 mL, 3.15 mmol) was added and thesolution was stirred at room temperature under argon atmosphere for 16h. It was then diluted with dichloromethane (40 mL) and washed with icecold aqueous citric acid (5 wt %, 30 mL) and water (2×20 mL). Theorganic phase was dried over anhydrous sodium sulfate and concentratedto dryness. The residue was used as such for the next step.

Cholesterol Derivatised CPG AI

Succinate AH (0.254 g, 0.242 mmol) was dissolved in a mixture ofdichloromethane/acetonitrile (3:2, 3 mL). To that solution DMAP (0.0296g, 0.242 mmol) in acetonitrile (1.25 mL),2,2′-Dithio-bis(5-nitropyridine) (0.075 g, 0.242 mmol) inacetonitrile/dichloroethane (3:1, 1.25 mL) were added successively. Tothe resulting solution triphenylphosphine (0.064 g, 0.242 mmol) inacetonitrile (0.6 ml) was added. The reaction mixture turned brightorange in color. The solution was agitated briefly using a wrist-actionshaker (5 mins). Long chain alkyl amine-CPG (LCAA-CPG) (1.5 g, 61 mM)was added. The suspension was agitated for 2 h. The CPG was filteredthrough a sintered funnel and washed with acetonitrile, dichloromethaneand ether successively. Unreacted amino groups were masked using aceticanhydride/pyridine. The achieved loading of the CPG was measured bytaking UV measurement (37 mM/g).

The synthesis of siRNAs bearing a 5′-12-dodecanoic acid bisdecylamidegroup (herein referred to as “5′-C32-”) or a 5′-cholesteryl derivativegroup (herein referred to as “5′-Chol-”) was performed as described inWO 2004/065601, except that, for the cholesteryl derivative, theoxidation step was performed using the Beaucage reagent in order tointroduce a phosphorothioate linkage at the 5′-end of the nucleic acidoligomer.

Nucleic acid sequences are represented below using standardnomenclature, and specifically the abbreviations of Table 1.

TABLE 1 Abbreviations of nucleotide monomers used in nucleic acidsequence representation. It will be understood that these monomers, whenpresent in an oligonucleotide, are mutually linked by5′-3′-phosphodiester bonds. Abbreviation Nucleotide(s) Aadenosine-5′-phosphate C cytidine-5′-phosphate G guanosine-5′-phosphateT 2′-deoxy-thymidine-5′-phosphate U uridine-5′-phosphate N anynucleotide (G, A, C, or T) a 2′-O-methyladenosine-5′-phosphate c2′-O-methylcytidine-5′-phosphate g 2′-O-methylguanosine-5′-phosphate u2′-O-methyluridine-5′-phosphate sT2′-deoxy-thymidine-5′phosphate-phosphorothioate

Example 2 Gene Walking of a Gene from the Ebola Virus

Design and in silico selection of siRNAs targeting Ebola virus

siRNA design was carried out to identify siRNAs targeting Ebola virusmRNAs for genes VP30, VP35, NP, L, VP24, VP40 and GP with a focus onsequences isolated from the Zaire region (EBOV-Z), as well as sequencesfrom Sudan (EBOV-S). The siRNA in silico selection resulted in 521siRNAs satisfying our selection criteria (Table 2).

Ebola Zaire sequence AY354458 was downloaded from NCBI Nucleotidedatabase and further on used as reference sequence for EBOV-Z. EbolaSudan sequence AY729654 was used as reference sequence for EBOV-S,respectively.

Sequence regions encoding target genes VP30, VP35, NP, L, VP24, VP40 andGP according to the information in the Genbank file were extracted fromboth reference sequences, followed by extraction of all possible 19mersfor each gene, resulting in the candidate siRNA target regions (andsiRNA sense sequences) for each distinct gene.

In order to identify siRNAs targeting all available EBOV-Z and EBOV-Ssequences, it was necessary to compile available Ebola sequences fromall sequenced isolates. For this, each of the 7 target gene sequencesextracted from the Ebola Zaire reference sequences was used in a blastsearch against viruses at NCBI with default parameters and resultingEbola mRNA hits were downloaded.

Each candidate target region was tested for conservation across theEbola sequences by searching the relevant target gene for presence ofthe 19mer target region. The percentage of conserved sequences acrossall downloaded sequences for the relevant gene was calculated for eachcandidate target region by dividing of the number of conserved sequenceswith the total number of downloaded sequences.

Example 3 Ebola siRNA In Vitro Screening

Table 3 provides a summary of the screening results of the siRNAsdescribed in Table 2. Following initial screening using a GFP-expressingEbola-Zaire virus and immunofluorescence screening using Ebola-Sudanvirus, siRNA showing activity were further tested by plaque assay foranti-viral activity against Ebola-Zaire and Ebola-Sudan strains. SeveralsiRNAs were identified that had significant activity against one or moreEbola strains. At a concentration of 100 nM many of the siRNA identifiedshowed greater than a 1 log reduction (>90% inhibition) in Ebola virustiters. Negative control luciferase and GFP siRNA at the sameconcentration showed reductions in virus titer of between 10 and 35%(Table 3). Three previously identified Ebola siRNA (LS L#1, LS NP#1, LSVP35#1) were also tested in parallel and these inhibited Ebola virus byroughly 70%. The previously identified Ebola siRNA are 25 nucleotideblunt-ended duplexes with the following composition: LS L#1, sense: 5′CCAUCUCUGAGACACGACAUAUCUU 3′ (SEQ ID NO: 1073) anti-sense: 5′AAGAUAUGUCGUGUCUCAGAGAUGG 3′ (SEQ ID NO: 1074); LS NP#1, sense: 5′GGUUCAAAGGCAAAUUCAAGUACAU 3′ (SEQ ID NO: 1075) anti-sense 5′AUGUACUUGAAUUUGCCUUUGAACC 3′ (SEQ ID NO: 1076); LS VP35#1, sense 5′CCCAAAUGCAACAAACCAAGCCAAA 3′ (SEQ ID NO: 1077) anti-sense 5′UUUGGCUUGGUUUGUUGCAUUUGGG 3′ (SEQ ID NO: 1078). The siRNA sequences forAD-1955 and AD-5179 are as follows: AD-1955, sense: 5′CUUACGCUGAGUACUUCGAdTsdT 3′ (SEQ ID NO: 1079) anti-sense: 5′UCGAAGUACUCAGCGUAAGdTsdT 3′ (SEQ ID NO: 1080); AD-5179, sense: 5′CcAcAuGAAGcAGcACGACusU 3′ (SEQ ID NO: 1081) anti-sense: 5′AAGUCGUGCUGCUUCAUGUGgsusC 3′ (SEQ ID NO: 1082).

Lead siRNAs were also screened for immunostimulatory activity (IFNalphaand TNFalpha). Immunostimulatory activity was assayed by transfectingsiRNAs into human peripheral blood mononuclear cells and measureingcytokine release by ELISA as outlined in Hornung et al. Nature Medicine2005. Cytokine levels were normalized to a positive control siRNAincluded in every assay. The lead candidates had no immunostimulatoryactivity.

The following procedures were used in generating the screening results.

GFP Ebola-Zaire Assay

VERO cells were transfected at ˜2X10E4 cells per well in a black-walled96 well plate. Transfection was performed in EMEM with 10% FCS overnightat 100, 10 and 1 nM siRNA complexed with lipofectamine (1.2 ul oflipofectamine per well in 50 ul volume; complexation was performed atroom temperature for 15-20 min).

Next day cells were infected with GFP-EBOLA virus (1/50 dilution ofstock EBOLA-Zaire GFP, stock E6(4) from 11OCT05, USAMRIID) in 50 ul ofEMEM with 10% FCS. 2 days later cells were fixed in 10% neutral-bufferedformalin for >3 days. Formalin was changed before removing from BSL-4suite. Next formalin was replaced with PBS.

To quantify infection level of cells in individual wells, cells werestained with 10-20 ul/well of 10 ug/ml Hoescht dye and read on Discovery1 microscope. GFP signal normalized to Hoescht signal was read as ameasure of infection level.

Immunofluorescence Ebola-Sudan Assay

VERO cells were transfected at ˜2X10E4 cell per well in a black-walled96 well plate. Transfection was performed in EMEM with 10% FCS overnightat 100, 10 and 1 nM siRNA complexed with lipofectamine (1.2 ul oflipofectamine per well in 50 ul volume; complexation was performed atroom temperature for 15-20 min).

Next day cells were infected with EBOLA-Sudan virus (1/100 dilution ofEBOV-Sudan (Boniface), stock GP(1)V(2)E6(2) from 23 May 06, USAMRIID) in50 μl of EMEM with 10% FCS. Two days later cells were fixed in 10%neutral-buffered formalin for >3 days. Formalin was changed beforeremoving from BSL-4 suite. Next formalin was replaced with PBS.

To detect infected cells, cells were stained for 4 h at room temperaturewith mouse anti-Sudan Boniface polyclonal sera (sera was collected from20 animals and pooled at day 30 post infection of C57BL/6 mice infectedwith ˜1000 pfu of the EBOLA SUDAN-BONIFACE, stock GP(1) V(2) V(1) E6(2)from 23 May 06, USAMRIID) at 1:200 dilution in PBS. Then cells werewashed with PBS 2× for 5 minutes. Goat anti-mouse IgG-AlexaFluor488(Molecular Probes) was added at 1:500 dilution in PBS. Cells were washedagain with PBS for 5 minutes, 100 ul of PBS was added to each well. Toquantify infection level of cells in individual wells, cell were stainedwith 10-20 ul/well of 10 ug/ml Hoescht dye and read on Discovery 1microscope. AlexaFluor488 signal normalized to Hoescht signal was readas a measure of infection level.

Plaque Assay for Filoviruses for In Vitro Assay

Vero cells were transfected in 24 well plates at the density of˜1.5X10E5/well density. Transfection was performed in EMEM with 10% FCSovernight at 100 nM siRNA complexed with lipofectamine (3 ul oflipofectamine per well in 200 ul volume; complexation was performed atroom temperature for 15-20 min). Transfection was done in duplicates. 24hours later duplicate plates were infected in 50 ul/well with either1/500 diluted Zaire-EBOV [(E6P2) stock from 20 Jun. 06, USAMRIID] or1/1000 diluted EBOV-Sudan [(strain Boniface), stock GP(1)V(2)E6(2) from23 May 06, USAMRIID]. After 1 hour at 37° C. virus inoculum was replacedwith 500 ul of fresh 10% FCS/EMEM.

48-72 h later supernatants were harvested from each well.

Plaque assay was performed with supernatants at 10⁻¹, 10⁻², 10⁻³, 10⁻⁴,10⁻⁵ and 10⁻⁶ dilutions. Fresh Vero cells in 6-well plates were infectedwith diluted supernatants for 1 hour at 37° C. with rocking plates every15 minutes; overlaid with 2 ml/well of 0.5% agarose in EMEM, 5% FCS,Pen/Strep. Six days later plates were overlaid with 2 ml of overlaymedia +4% neutral red solution and read plates the following day.

siRNA Activity Determination Using the Plasmid System.

Consensus sequences of NP (SEQ ID NO:1043), GP (SEQ ID NO:1044), L, VP24(SEQ ID NO:1045), VP30 (SEQ ID NO:1046), VP35 (SEQ ID NO:1047), VP40(SEQ ID NO:1048) were synthesized by GENEART (Regensburg, Germany) andcloned into GENEART standard vector. The L gene was generated as 2fragments (SEQ ID NO:1049 and SEQ ID NO:1050). All genes were subclonedinto individual psiCheck-2 (Promega, Mannheim, Germany) vectors via XhoIand NotI sites, resulting in a construct with the flu gene between thestop-codon and the polyA-signal of Renilla luciferase. Correct cloningwas confirmed by end sequencing performed by GATC Biotech (Konstanz,Germany).

Transfections:

Cos-7 cells were seeded at 1.5×10⁴ cells/well on white 96-well plateswith clear bottom (Greiner Bio-One GmbH, Frickenhausen, Germany) in 75μl of growth medium. Directly after seeding the cells, 50 ng ofplasmid/well were transfected with Lipofectamine-2000 (Invitrogen) asdescribed below for the siRNAs, with the plasmid diluted in Opti-MEM toa final volume of 12.5 μl/well, prepared as a mastermix for the wholeplate.

siRNA transfections were performed in quadruplicates 4 h after plasmidtransfection. For each well 0.5 μl Lipofectamine-2000 (Invitrogen GmbH,Karlsruhe, Germany) were mixed with 12 μl Opti-MEM (Invitrogen) andincubated for 15 min at room temperature. For an siRNA concentration of50 nM in the 100 μl transfection volume, 1 μl of a 5 μM siRNA were mixedwith 11.5 μl Opti-MEM per well, combined with theLipofectamine-2000-Opti-MEM mixture and again incubated for 15 minutesat room temperature. During incubation, the growth medium was removedfrom cells and replaced by 75 μl/well of fresh medium.siRNA-Lipofectamine2000-complexes were applied completely (25 μl eachper well) to the cells and cells were incubated for 24 h at 37° C. and5% CO₂ in a humidified incubator (Heraeus GmbH, Hanau, Germany).

Cells were harvested by removing growth medium and application of 150 μlof a 1:1 mixture consisting of medium and Dual-Glo Luciferase substrate,from the Dual-Glo Luciferase Assay System (Promega, Mannheim, Germany).The luciferase assay was performed according to the manufacturer'sprotocol for Dual-Glo Luciferase assay and luminescence was measured ina Victor-Light 1420 Luminescence Counter (Perkin Elmer,Rodgau-Jügesheim, Germany). Values obtained with Renilla luciferase werenormalized to the respective values obtained with Firefly luciferase.Values acquired with siRNAs directed against an Ebola gene werenormalized to the value obtained with an unspecific siRNA (directedagainst neomycin resistance gene) set to 100%.

Example 4 In Vivo Filovirus Infection Model

Liposome-formulated siRNAs targeting Ebola genes protected mice from alethal Ebola virus challenge. Details on the liposome formulation aredetailed in the next section. Mice were treated with liposome-formulatedsiRNA (described below) twice, at 2 hours prior to Ebola infection (5mg/kg i.v.) and at 3 days after Ebola infection (3 mg/kg i.p.). Micewere infected intraperitoneally with 30,000 LD50 of Ebola-Zaire (LD50 islethal dose of Ebola infection where 50% of animals die). Mice weremonitored for survival with n=10 per treatment group. Negative controlsincluded untreated mice and mice treated with liposome-formulatedluciferase siRNA (AD-1955). EK1 is a previously published siRNA sequencetargeting the Ebola L gene [Geisbert et al. (2006) The Journal ofInfectious Disease 193:1950-1657] that was used as a positive control.The siRNA sequences for EK1 are as follows:

(SEQ ID NO: 1083) 5′ GUACGAAGCUGUAUAUAAAdTdT 3′ (sense) and(SEQ ID NO: 1084) 5′ UUUAUAUACAGCUUCGUACdTdT 3′ (antisense).

FIG. 1 provides the results. All the negative control-treated animals(untreated and liposomally-formulated luciferase siRNA-treated) diedwithin 6-8 days following Ebola infection. Several of theliposomally-formulated Ebola siRNAs showed significant increases insurvival rates compared to the negative controls. Multiple Ebola siRNA(AD-11691, AD-11710, AD-11588, AD-11599, AD-11570) showed moreprotection against lethal Ebola infection than the previously publishedEK1 siRNA (FIG. 1).

A further experiment was conducted utilizing one of the active EbolasiRNA (AD-11570) to investigate different dosing routes and treatmentregimens. Mice were treated with liposome-formulated Ebola VP35 siRNA(AD-11570) or negative control luciferase siRNA (AD-1955) 2 hours priorto Ebola infection (5 mg/kg i.p.). Mice were infected intraperitoneallywith 30,000 LD50 of Ebola-Zaire (LD50 is lethal dose of Ebola infectionwhere 50% of animals die). Mice were monitored for survival with n=10per treatment group.

FIG. 2 provides the results. The animals treated withlipsomally-formulated AD-11570 showed near complete protection againstlethal Ebola infection as compared to the negative control-treatedanimals (untreated and liposomally-formulated luciferase siRNA-treated)(FIG. 2). These results indicate that a single siRNA administrationeither via intravenous or intraperitoneal route is able to have asignificant impact on survival.

Formulation Procedure

The lipidoid ND98.4HCl (MW 1487) (FIG. 3), Cholesterol (Sigma-Aldrich),and PEG-Ceramide C16 (Avanti Polar Lipids) were used to preparelipid-siRNA nanoparticles. Stock solutions of each in ethanol wereprepared: ND98, 133 mg/mL; Cholesterol, 25 mg/mL, PEG-Ceramide C16, 100mg/mL. ND98, Cholesterol, and PEG-Ceramide C16 stock solutions were thencombined in a 42:48:10 molar ratio. Combined lipid solution was mixedrapidly with aqueous siRNA (in sodium acetate pH 5) such that the finalethanol concentration was 35-45% and the final sodium acetateconcentration was 100-300 mM. Lipid-siRNA nanoparticles formedspontaneously upon mixing. Depending on the desired particle sizedistribution, the resultant nanoparticle mixture was in some casesextruded through a polycarbonate membrane (100 nm cut-off) using athermobarrel extruder (Lipex Extruder, Northern Lipids, Inc). In othercases, the extrusion step was omitted. Ethanol removal and simultaneousbuffer exchange was accomplished by either dialysis or tangential flowfiltration. Buffer was exchanged to phosphate buffered saline (PBS) pH7.2.

Characterization of Formulations

Formulations prepared by either the standard or extrusion-free methodare characterized in a similar manner. Formulations are firstcharacterized by visual inspection. They should be whitish translucentsolutions free from aggregates or sediment. Particle size and particlesize distribution of lipid-nanoparticles are measured by dynamic lightscattering using a Malvern Zetasizer Nano ZS (Malvern, USA). Particlesshould be 20-300 nm, and ideally, 40-100 nm in size. The particle sizedistribution should be unimodal. The total siRNA concentration in theformulation, as well as the entrapped fraction, is estimated using a dyeexclusion assay. A sample of the formulated siRNA is incubated with theRNA-binding dye Ribogreen (Molecular Probes) in the presence or absenceof a formulation disrupting surfactant, 0.5% Triton-X100. The totalsiRNA in the formulation is determined by the signal from the samplecontaining the surfactant, relative to a standard curve. The entrappedfraction is determined by subtracting the “free” siRNA content (asmeasured by the signal in the absence of surfactant) from the totalsiRNA content. Percent entrapped siRNA is typically >85%.

Example 5 siRNAs Targeting Ebola Increased the Life-Span of Mice andGuinea Pigs Infected with Ebola

siRNA directed against different Ebola genes were formulated inliposomes. A single dose of siRNA targeting the VP35 gene (AD-11570)protected both mice and guinea pigs against lethal Ebola infection (1000PFU; 30000x LD50). Protection was associated with reduction in viraltitres and was seen when drug was administered either prophylacticallyor therapeutically. Irrelevant siRNA (targeting luciferase) similarlyformulated showed no protective effect or impact on virus titer.

Studies were conducted in mouse, guinea pig, and nonhuman primate lethalEbola infection models. The studies are summarized below.

Mouse study 1: Demonstrated efficacy in mouse model of Ebola formultiple siRNA sequences formulated in LNP01. siRNAs were administeredby intravenous (i.v.) injection on day 0, followed by intraperitoneal(i.p.) injection on day 3. See FIG. 1.

Mouse study 3: Demonstrated efficacy of siRNA in LNP01 formulation inthe mouse model of Ebola when given by either IV or IP route. See FIG.2. Mice were monitored for survival with n=10 per treatment group.

Mouse study #14: Demonstrated efficacy of siRNA in DODMA in the mousemodel of Ebola by the IP route. See FIG. 4. siRNAs were formulated inDODMA:DSPC:Chol:PEG-DMG. Mice were monitored for survival with n=10 pertreatment group. Treatment with 10 mg/kg DODMA-formulated AD-11570siRNAs was also effective to protect guinea pigs infected with Ebola.

Mouse study #15: Demonstrated that siRNA in DODMA formulation iseffective down to 0.04 mg/kg in the mouse model of Ebola. AD-11570consistently gave 25-50% protection, but no clear dose response wasseen. The control siRNA AD-1955 gave 25-50% protection, but again, nodose response was observed. See FIG. 5.

Guinea pig study #6: Demonstrated efficacy of multiple doses of siRNA inDODMA formulation in the guinea pig model of Ebola. AD-11570 siRNAsformulated in DODMA:DSPC:Chol:PEG-DMG were effective to protect guineapigs from Ebola. See FIG. 6.

Guinea pig study #11: Efficacy of siRNAs formulated with DODMA andtargeting different Ebola genes in a guinea pig model of Ebola. See FIG.7

A 95% decrease in viral titers was also observed followingadministration of LNP01 formulated VP35 siRNA to BALB/c mice (n=5 pergroup) (FIG. 8). Mice were dosed systemically with LNP01 formulatedsiRNA at 5 mg/kg i.v. at day 0, then 3 mg/kg i.p. at day 3. Two hourspost-siRNA injection at day 0, mice were injected with 1,000 pfuEbola-Zaire virus and monitored for survival. On day 6 post-infection,the mice were sacrificed and their blood viral titers were determined byplaque assay.

Table 3 shows the results of cell-based and plaque assays.

Table 4 shows the results of plaque assays for control siRNAs.

Table 5. shows the sequences of modified duplexes, and Table 6 shows theeffect of the modified duplexes on plaque assay activity and IC₅₀ valuesin the plasmid-based system.

Table 7 shows the effect of siRNAs on cytokine levels (IFN-alpha andTNF-alpha).

Table 8 shows the siRNA silencing in the plasmid system and calculatedIC₅₀ values.

Table 9 shows that nonhuman primates administered siRNAs targeting Eboladid not demonstrate a decrease in lymphocyte or platelet count.

TABLE 2 position double overhang design antisense strand in sense strandSeq duplex target Target name Seq ID sequence (5′-3′) name IDsequence (5′-3′) name    3-21 VP35 A18480 1 GAUGAAGAUUAAAACCUUCTsTA18481 2 GAAGGUUUUAAUCUUCAUCTsT AD- 11542    4-22 VP35 A18482 3AUGAAGAUUAAAACCUUCATsT A18483 4 UGAAGGUUUUAAUCUUCAUTsT AD- 11543    5-23VP35 A18484 5 UGAAGAUUAAAACCUUCAUTsT A18485 6 AUGAAGGUUUUAAUCUUCATsT AD-11544    6-24 VP35 A18486 7 GAAGAUUAAAACCUUCAUCTsT A18487 8GAUGAAGGUUUUAAUCUUCTsT AD- 11545 1354- VP35 A18488 9UGAUGAAGAUUAAGAAAAATsT A18489 10 UUUUUCUUAAUCUUCAUCATsT AD- 1372 11546   7-25 VP35 A18490 11 AAGAUUAAAACCUUCAUCATsT A18491 12UGAUGAAGGUUUUAAUCUUTsT AD- 11547    8-26 VP35 A18492 13AGAUUAAAACCUUCAUCAUTsT A18493 14 AUGAUGAAGGUUUUAAUCUTsT AD- 11548   9-27 VP35 A18494 15 GAUUAAAACCUUCAUCAUCTsT A18495 16GAUGAUGAAGGUUUUAAUCTsT AD- 11549   10-28 VP35 A18496 17AUUAAAACCUUCAUCAUCCTsT A18497 18 GGAUGAUGAAGGUUUUAAUTsT AD- 11550  11-29 VP35 A18498 19 UUAAAACCUUCAUCAUCCUTsT A18499 20AGGAUGAUGAAGGUUUUAATsT AD- 11551   12-30 VP35 A18500 21UAAAACCUUCAUCAUCCUUTsT A18501 22 AAGGAUGAUGAAGGUUUUATsT AD- 11552   1-19 VP30 A18502 23 GAUGAAGAUUAAGAAAAAGTsT A18503 24CUUUUUCUUAAUCUUCAUCTsT AD- 11553    3-21 VP35 A18504 151GAuGAAGAuuAAAAccuucTsT A18505 152 GAAGGUUUuAAUCUUcAUCTsT AD- 11554   4-22 VP35 A18506 153 AuGAAGAuuAAAAccuucATsT A18507 154UGAAGGUUUuAAUCUUcAUTsT AD- 11555    5-23 VP35 A18508 155uGAAGAuuAAAAccuucAuTsT A18509 156 AUGAAGGUUUuAAUCUUcATsT AD- 11556   6-24 VP35 A18510 157 GAAGAuuAAAAccuucAucTsT A18511 158GAUGAAGGUUUuAAUCUUCTsT AD- 11557 1354- VP35 A18512 159uGAuGAAGAuuAAGAAAAATsT A18513 160 UUUUUCUuAAUCUUcAUcATsT AD- 1372 11558   7-25 VP35 A18514 161 AAGAuuAAAAccuucAucATsT A18515 162UGAUGAAGGUUUuAAUCUUTsT AD- 11559    8-26 VP35 A18516 163AGAuuAAAAccuucAucAuTsT A18517 164 AUGAUGAAGGUUUuAAUCUTsT AD- 11560   9-27 VP35 A18518 165 GAuuAAAAccuucAucAucTsT A18519 166GAUGAUGAAGGUUUuAAUCTsT AD- 11561   10-28 VP35 A18520 167AuuAAAAccuucAucAuccTsT A18521 168 GGAUGAUGAAGGUUUuAAUTsT AD- 11562  11-29 VP35 A18522 169 uuAAAAccuucAucAuccuTsT A18523 170AGGAUGAUGAAGGUUUuAATsT AD- 11563   12-30 VP35 A18524 171uAAAAccuucAucAuccuuTsT A18525 172 AAGGAUGAUGAAGGUUUuATsT AD- 11564   1-19 VP30 A18526 173 GAuGAAGAuuAAGAAAAAGTsT A18527 174CUUUUUCUuAAUCUUcAUCTsT AD- 11565    3-21 VP35 A18528 1019GAuGAAGAuuAAAAccuucTsT A18529 1020 GAAGGuuuUAAUCuUcAUCTsT AD- 11566   4-22 VP35 A18530 1021 AuGAAGAuuAAAAccuucATsT A18531 1022uGAAGGuuuUAAUCuUcAUTsT AD- 11567    5-23 VP35 A18532 1023uGAAGAuuAAAAccuucAuTsT A18533 1024 AuGAAGGuuuuAAUCuUcATsT AD- 11568   6-24 VP35 A18534 1025 GAAGAuuAAAAccuucAucTsT A18535 1026GAuGAAGGuuuuAAUCuUCTsT AD- 11569 1354- VP35 A18536 1027uGAuGAAGAuuAAGAAAAATsT A18537 1028 uuuuUCuuAAUCuUcAUcATsT AD- 1372 11570   7-25 VP35 A18538 1029 AAGAuuAAAAccuucAucATsT A18539 1030uGAuGAAGGuuuuAAUCuUTsT AD- 11571    8-26 VP35 A18540 1031AGAuuAAAAccuucAucAuTsT A18541 1032 AuGAuGAAGGuuuuAAUCUTsT AD- 11572   9-27 VP35 A18542 1033 GAuuAAAAccuucAucAucTsT A18543 1034GAuGAuGAAGGuuuuAAUCTsT AD- 11573   10-28 VP35 A18544 1035AuuAAAAccuucAucAuccTsT A18545 1036 GGAuGAuGAAGGuuuuAAUTsT AD- 11574  11-29 VP35 A18546 1037 uuAAAAccuucAucAuccuTsT A18547 1038AGGAuGAuGAAGGuuuuAATsT AD- 11575   12-30 VP35 A18548 1039uAAAAccuucAucAuccuuTsT A18549 1040 AAGGAuGAuGAAGGuuuuATsT AD- 11576   1-19 VP30 A18550 1041 GAuGAAGAuuAAGAAAAAGTsT A18551 1042CuuuuUCuuAAUCuUcAUCTsT AD- 11577  996-1014 NP A18552 25UGGACACAUGAUGGUGAUCTsT A18553 26 GAUCACCAUCAUGUGUCCATsT AD- 11578 1467-NP A18554 27 AAGCAACUCCAACAAUAUGTsT A18555 28 CAUAUUGUUGGAGUUGCUUTsT AD-1485 11579   11-29 VP35 A18556 29 AAAACCUUCAUCAUCCUUUTsT A18557 30AAAGGAUGAUGAAGGUUUUTsT AD- 11580 1357- VP35 A18558 31UUGAUGAAGAUUAAGAAAATsT A18559 32 UUUUCUUAAUCUUCAUCAATsT AD- 1375 11581   1-19 VP40 A18560 33 GAUGAAGAUUAAGAAAAAGTsT A18561 34CUUUUUCUUAAUCUUCAUCTsT AD- 11582  647-665 VP40 A18562 35CCCUGCUGCAACAUGGACATsT A18563 36 UGUCCAUGUUGCAGCAGGGTsT AD- 11583   2-20 VP30 A18564 37 AUGAAGAUUAAGAAAAAGUTsT A18565 38ACUUUUUCUUAAUCUUCAUTsT AD- 11584    1-19 L A18566 39AAGAUUAAGAAAAAGUCCATsT A18567 40 UGGACUUUUUCUUAAUCUUTsT AD- 11585   5-23 NP A18568 41 AAGAUUAAUAAUUUUCCUCTsT A18569 42GAGGAAAAUUAUUAAUCUUTsT AD- 11586  823-841 NP A18570 43AUGCCGGAAGAGGAGACAATsT A18571 44 UUGUCUCCUCUUCCGGCAUTsT AD- 11587 824-842 NP A18572 45 UGCCGGAAGAGGAGACAACTsT A18573 46GUUGUCUCCUCUUCCGGCATsT AD- 11588  987-1005 NP A18574 47GCAAUCAGUAGGACACAUGTsT A18575 48 CAUGUGUCCUACUGAUUGCTsT AD- 11589 988-1006 NP A18576 49 CAAUCAGUAGGACACAUGATsT A18577 50UCAUGUGUCCUACUGAUUGTsT AD- 11590  989-1007 NP A18578 51AAUCAGUAGGACACAUGAUTsT A18579 52 AUCAUGUGUCCUACUGAUUTsT AD- 11591 990-1008 NP A18580 53 AUCAGUAGGACACAUGAUGTsT A18581 54CAUCAUGUGUCCUACUGAUTsT AD- 11592  991-1009 NP A18582 55UCAGUAGGACACAUGAUGGTsT A18583 56 CCAUCAUGUGUCCUACUGATsT AD- 11593 992-1010 NP A18584 57 CAGUAGGACACAUGAUGGUTsT A18585 58ACCAUCAUGUGUCCUACUGTsT AD- 11594  993-1011 NP A18586 59AGUAGGACACAUGAUGGUGTsT A18587 60 CACCAUCAUGUGUCCUACUTsT AD- 11595 994-1012 NP A18588 61 GUAGGACACAUGAUGGUGATsT A18589 62UCACCAUCAUGUGUCCUACTsT AD- 11596  995-1013 NP A18590 63UAGGACACAUGAUGGUGAUTsT A18591 64 AUCACCAUCAUGUGUCCUATsT AD- 11597 1005-NP A18592 65 GAUGGUGAUUUUCCGUUUGTsT A18593 66 CAAACGGAAAAUCACCAUCTsT AD-1023 11598 1006- NP A18594 67 AUGGUGAUUUUCCGUUUGATsT A18595 68UCAAACGGAAAAUCACCAUTsT AD- 1024 11599 1007- NP A18596 69UGGUGAUUUUCCGUUUGAUTsT A18597 70 AUCAAACGGAAAAUCACCATsT AD- 1025 116001008- NP A18598 71 GGUGAUUUUCCGUUUGAUGTsT A18599 72CAUCAAACGGAAAAUCACCTsT AD- 1026 11601 1462- NP A18600 73GCUGAGAAGCAACUCCAACTsT A18601 74 GUUGGAGUUGCUUCUCAGCTsT AD- 1480 116021463- NP A18602 75 CUGAGAAGCAACUCCAACATsT A18603 76UGUUGGAGUUGCUUCUCAGTsT AD- 1481 11603 1464- NP A18604 77UGAGAAGCAACUCCAACAATsT A18605 78 UUGUUGGAGUUGCUUCUCATsT AD- 1482 116041465- NP A18606 79 GAGAAGCAACUCCAACAAUTsT A18607 80AUUGUUGGAGUUGCUUCUCTsT AD- 1483 11605 1466- NP A18608 81AGAAGCAACUCCAACAAUATsT A18609 82 UAUUGUUGGAGUUGCUUCUTsT AD- 1484 116061353- VP35 A18610 83 AAAAGUGAUGAAGAUUAAGTsT A18611 84CUUAAUCUUCAUCACUUUUTsT AD- 1371 11607 1354- VP35 A18612 85AAAGUGAUGAAGAUUAAGATsT A18613 86 UCUUAAUCUUCAUCACUUUTsT AD- 1372 116081355- VP35 A18614 87 AAGUGAUGAAGAUUAAGAATsT A18615 88UUCUUAAUCUUCAUCACUUTsT AD- 1373 11609 1356- VP35 A18616 89AGUGAUGAAGAUUAAGAAATsT A18617 90 UUUCUUAAUCUUCAUCACUTsT AD- 1374 11610 645-663 VP40 A18618 91 CUGCCUGCUGCAACAUGGATsT A18619 92UCCAUGUUGCAGCAGGCAGTsT AD- 11611  646-664 VP40 A18620 93UGCCUGCUGCAACAUGGACTsT A18621 94 GUCCAUGUUGCAGCAGGCATsT AD- 11612 451-469 GP A18622 95 GGCUGAAAACUGCUACAAUTsT A18623 96AUUGUAGCAGUUUUCAGCCTsT AD- 11613  452-470 GP A18624 97GCUGAAAACUGCUACAAUCTsT A18625 98 GAUUGUAGCAGUUUUCAGCTsT AD- 11614 453-471 GP A18626 99 CUGAAAACUGCUACAAUCUTsT A18627 100AGAUUGUAGCAGUUUUCAGTsT AD- 11615  454-472 GP A18628 101UGAAAACUGCUACAAUCUUTsT A18629 102 AAGAUUGUAGCAGUUUUCATsT AD- 11616 455-473 GP A18630 103 GAAAACUGCUACAAUCUUGTsT A18631 104CAAGAUUGUAGCAGUUUUCTsT AD- 11617  456-474 GP A18632 105AAAACUGCUACAAUCUUGATsT A18633 106 UCAAGAUUGUAGCAGUUUUTsT AD- 11618 457-475 GP A18634 107 AAACUGCUACAAUCUUGAATsT A18635 108UUCAAGAUUGUAGCAGUUUTsT AD- 11619  458-476 GP A18636 109AACUGCUACAAUCUUGAAATsT A18637 110 UUUCAAGAUUGUAGCAGUUTsT AD- 11620 459-477 GP A18638 111 ACUGCUACAAUCUUGAAAUTsT A18639 112AUUUCAAGAUUGUAGCAGUTsT AD- 11621  599-617 VP30 A18640 113AGCAAAUCCAACGGCUGAUTsT A18641 114 AUCAGCCGUUGGAUUUGCUTsT AD- 11622 600-618 VP30 A18642 115 GCAAAUCCAACGGCUGAUGTsT A18643 116CAUCAGCCGUUGGAUUUGCTsT AD- 11623  601-619 VP30 A18644 117CAAAUCCAACGGCUGAUGATsT A18645 118 UCAUCAGCCGUUGGAUUUGTsT AD- 11624 135-153 L A18646 119 UUGGACCAAUGUGACCUAGTsT A18647 120CUAGGUCACAUUGGUCCAATsT AD- 11625  136-154 L A18648 121UGGACCAAUGUGACCUAGUTsT A18649 122 ACUAGGUCACAUUGGUCCATsT AD- 11626 2100-L A18650 123 AUGCAUGUCAGUGAUUAUUTsT A18651 124 AAUAAUCACUGACAUGCAUTsTAD- 2118 11627 2101- L A18652 125 UGCAUGUCAGUGAUUAUUATsT A18653 126UAAUAAUCACUGACAUGCATsT AD- 2119 11628 2102- L A18654 127GCAUGUCAGUGAUUAUUAUTsT A18655 128 AUAAUAAUCACUGACAUGCTsT AD- 2120 116292103- L A18656 129 CAUGUCAGUGAUUAUUAUATsT A18657 130UAUAAUAAUCACUGACAUGTsT AD- 2121 11630 2104- L A18658 131AUGUCAGUGAUUAUUAUAATsT A18659 132 UUAUAAUAAUCACUGACAUTsT AD- 2122 116312114- L A18660 133 UUAUUAUAAUCCACCACAUTsT A18661 134AUGUGGUGGAUUAUAAUAATsT AD- 2132 11632 2115- L A18662 135UAUUAUAAUCCACCACAUATsT A18663 136 UAUGUGGUGGAUUAUAAUATsT AD- 2133 116332116- L A18664 137 AUUAUAAUCCACCACAUAATsT A18665 138UUAUGUGGUGGAUUAUAAUTsT AD- 2134 11634 2412- L A18666 139AAAGUUACAAGUGCCUGUGTsT A18667 140 CACAGGCACUUGUAACUUUTsT AD- 2430 116352413- L A18668 141 AAGUUACAAGUGCCUGUGGTsT A18669 142CCACAGGCACUUGUAACUUTsT AD- 2431 11636 2466- L A18670 143UCAGGUUUUAUCUAUUUUGTsT A18671 144 CAAAAUAGAUAAAACCUGATsT AD- 2484 116372467- L A18672 145 CAGGUUUUAUCUAUUUUGGTsT A18673 146CCAAAAUAGAUAAAACCUGTsT AD- 2485 11638 2556- L A18674 147UCUGAUGCAAUUUUUGAUGTsT A18675 148 CAUCAAAAAUUGCAUCAGATsT AD- 2574 116392557- L A18676 149 CUGAUGCAAUUUUUGAUGATsT A18677 150UCAUCAAAAAUUGCAUCAGTsT AD- 2575 11640 1825- NP A18678 1085AGuuAcucGGAAAAcGGcATsT A18679 1086 UGCCGUUUUCCGAGuAACUTsT AD- 1843 116411588- NP A18680 1087 AAcGcuAuGGuAAcucuAATsT A18681 1088UuAGAGUuACcAuAGCGUUTsT AD- 1606 11642 1827- NP A18682 1089uuAcucGGAAAAcGGcAuGTsT A18683 1090 cAUGCCGUUUUCCGAGuAATsT AD- 1845 116431583- NP A18684 1091 AAAcAAAcGcuAuGGuAAcTsT A18685 1092GUuACcAuAGCGUUUGUUUTsT AD- 1601 11644 1488- NP A18686 1093AGAGucucGcGAAcuuGAcTsT A18687 1094 GUcAAGUUCGCGAGACUCUTsT AD- 1506 116451489- NP A18688 1095 GAGucucGcGAAcuuGAccTsT A18689 1096GGUcAAGUUCGCGAGACUCTsT AD- 1507 11646 1585- NP A18690 1097AcAAAcGcuAuGGuAAcucTsT A18691 1098 GAGUuACcAuAGCGUUUGUTsT AD- 1603 116471586- NP A18692 1099 cAAAcGcuAuGGuAAcucuTsT A18693 1100AGAGUuACcAuAGCGUUUGTsT AD- 1604 11648 2231- NP A18694 1101cAccGGcucccGuAuAcAGTsT A18695 1102 CUGuAuACGGGAGCCGGUGTsT AD- 2249 116492873- NP A18696 1103 cuAAcuAGcGAuuuAucuATsT A18697 1104uAGAuAAAUCGCuAGUuAGTsT AD- 2891 11650 1172- VP35 A18698 1105GcuGAAcuAuAGGGuAcGuTsT A18699 1106 ACGuACCCuAuAGUUcAGCTsT AD- 1190 116511176- VP35 A18700 1107 AAcuAuAGGGuAcGuuAcATsT A18701 1108UGuAACGuACCCuAuAGUUTsT AD- 1194 11652 1174- VP35 A18702 175uGAAcuAuAGGGuAcGuuATsT A18703 176 uAACGuACCCuAuAGUUcATsT AD- 1192 116531178- VP35 A18706 177 cuAuAGGGuAcGuuAcAuuTsT A18707 178AAUGuAACGuACCCuAuAGTsT AD- 1196 11655  251-269 VP35 A18704 179GGAuuAuGcuAcGcAucccTsT A18705 180 GGGAUGCGuAGcAuAAUCCTsT AD- 11654 416-434 VP35 A18708 181 uuAGAAcAAcGcAuuAcGATsT A18709 182UCGuAAUGCGUUGUUCuAATsT AD- 11656  421-439 VP35 A18710 183AcAAcGcAuuAcGAGucuuTsT A18711 184 AAGACUCGuAAUGCGUUGUTsT AD- 11657 1057-VP35 A18712 185 uGAucGAGGuuGGGuAuGuTsT A18713 186 AcAuACCcAACCUCGAUcATsTAD- 1075 11658  167-185 GP A18714 187 ccucGuGAucGAuucAAGATsT A18715 188UCUUGAAUCGAUcACGAGGTsT AD- 11659  163-181 GP A18716 189GuuAccucGuGAucGAuucTsT A18717 190 GAAUCGAUcACGAGGuAACTsT AD- 11660 658-676 GP A18720 191 AAcGAcuuucGcuGAAGGuTsT A18721 192ACCUUcAGCGAAAGUCGUUTsT AD- 11662  755-773 GP A18722 193AcGGAGGAcccGucuAGuGTsT A18723 194 cACuAGACGGGUCCUCCGUTsT AD- 11663 966-984 GP A18724 195 AGGucAAccccGAAAuuGATsT A18725 196UcAAUUUCGGGGUUGACCUTsT AD- 11664  978-996 GP A18726 197AAAuuGAuAcAAcAAucGGTsT A18727 198 CCGAUUGUUGuAUcAAUUUTsT AD- 11665 985-1003 GP A18728 199 uAcAAcAAucGGGGAGuGGTsT A18729 200CcACUCCCCGAUUGUUGuATsT AD- 11666 1101- GP A18730 201AGAGuccGGcGcGAAcuucTsT A18731 202 GAAGuUCGCGCCGGACUCUTsT AD- 1119 116671730- GP A18718 203 uGGAuAccAuAuuucGGGcTsT A18719 204GCCCGAAAuAUGGuAUCcATsT AD- 1748 11661 1820- GP A18732 205cuGGccAAcGAGAcGAcucTsT A18733 206 GAGUCGUCUCGUUGGCcAGTsT AD- 1838 116681298- VP30 A18734 207 uAucGcucGuAAuAuAAccTsT A18735 208GGUuAuAUuACGAGCGAuATsT AD- 1316 11669  295-313 VP30 A18736 209uucGAGcAcGAucAucAucTsT A18737 210 GAuGAuGAUCGuGCUCGAATsT AD- 11670 590-608 VP30 A18738 211 cucGcGcuuAGcAAAuccATsT A18739 212UGGAUUUGCuAAGCGCGAGTsT AD- 11671  519-537 VP30 A18740 213uuAcuccuAcuAAucGcccTsT A18741 214 GGGCGAUuAGuAGGAGuAATsT AD- 11672 126-144 VP30 A18742 215 cuGcGAAccGGuAGAGuuuTsT A18743 216AAACUCuACCGGUUCGcAGTsT AD- 11673  133-151 VP30 A18744 217ccGGuAGAGuuuAGuuGcATsT A18745 218 UGcAACuAAACUCuACCGGTsT AD- 11674 292-310 VP30 A18746 219 AuGuucGAGcAcGAucAucTsT A18747 220GAUGAUCGUGCUCGAAcAUTsT AD- 11675  321-339 VP30 A18748 221AAuuAucGAGGuGAGuAccTsT A18749 222 GGuACUcACCUCGAuAAUUTsT AD- 11676 910-928 VP30 A18750 223 GGGAccGAcAAucccuAAuTsT A18751 224AUuAGGGAUUGUCGGUCCCTsT AD- 11677 1295- VP30 A18752 225ucGuAucGcucGuAAuAuATsT A18753 226 uAuAUuACGAGCGAuACGATsT AD- 1313 11678 331-349 VP30 A18754 227 GuGAGuAccGucAAucAAGTsT A18755 228CUUGAUUGACGGuACUcACTsT AD- 11679  123-141 VP30 A18756 229GAucuGcGAAccGGuAGAGTsT A18757 230 CUCuACCGGUUCGcAGAUCTsT AD- 11680 124-142 VP30 A18758 231 AucuGcGAAccGGuAGAGuTsT A18759 232ACUCuACCGGUUCGcAGAUTsT AD- 11681 1293- VP30 A18760 233ucucGuAucGcucGuAAuATsT A18761 234 uAUuACGAGCGAuACGAGATsT AD- 1311 11682 145-163 VP30 A18762 235 AGuuGcAAccuAAcAcAcATsT A18763 236UGUGUGUuAGGUUGcAACUTsT AD- 11683  293-311 VP30 A18764 237uGuucGAGcAcGAucAucATsT A18765 238 UGAUGAUCGUGCUCGAAcATsT AD- 11684 358-376 VP30 A18766 239 cAcAAGuGcGcGuuccuAcTsT A18767 240GuAGGAACGCGcACUUGUGTsT AD- 11685  359-377 VP30 A18768 241AcAAGuGcGcGuuccuAcuTsT A18769 242 AGuAGGAACGCGcACUUGUTsT AD- 11686 518-536 VP30 A18770 243 AuuAcuccuAcuAAucGccTsT A18771 244GGCGAUuAGuAGGAGuAAUTsT AD- 11687  520-538 VP30 A18772 245uAcuccuAcuAAucGcccGTsT A18773 246 CGGGCGAUuAGuAGGAGuATsT AD- 11688 524-542 VP30 A18774 247 ccuAcuAAucGcccGuAAGTsT A18775 248CUuACGGGCGAUuAGuAGGTsT AD- 11689  525-543 VP30 A18776 249cuAcuAAucGcccGuAAGATsT A18777 250 UCUuACGGGCGAUuAGuAGTsT AD- 11690 584-602 VP30 A18778 251 cAAGGAcucGcGcuuAGcATsT A18779 252UGCuAAGCGCGAGUCCUUGTsT AD- 11691  469-487 VP24 A18784 253AGcuAcGGGAcGAuAcAAuTsT A18785 254 AUUGuAUCGUCCCGuAGCUTsT AD- 11694 910-928 VP24 A18786 255 GucGuuGAuucGAuccAAuTsT A18787 256AUUGGAUCGAAUcAACGACTsT AD- 11695  467-485 VP24 A18788 257AAAGcuAcGGGAcGAuAcATsT A18789 258 UGuAUCGUCCCGuAGCUUUTsT AD- 11696 862-880 VP24 A18792 259 cAAcAuGcGAAcAcAAcGuTsT A18793 260ACGUUGUGUUCGcAUGUUGTsT AD- 11698  466-484 VP24 A18796 261uAAAGcuAcGGGAcGAuAcTsT A18797 262 GuAUCGUCCCGuAGCUUuATsT AD- 11700 523-541 VP24 A18804 263 uGucuuAAGcGAccucuGuTsT A18805 264AcAGAGGUCGCUuAAGAcATsT AD- 11704  958-976 VP24 A18806 265ucuAcAuGucGuGAAcuAcTsT A18807 266 GuAGUUcACGAcAUGuAGATsT AD- 11705 959-977 VP24 A18808 267 cuAcAuGucGuGAAcuAcATsT A18809 268UGuAGUUcACGAcAUGuAGTsT AD- 11706  971-989 VP24 A18810 269AAcuAcAAcGGAuuGuuGATsT A18811 270 UcAAcAAUCCGUUGuAGUUTsT AD- 11707 1071-VP24 A18812 271 ccGAcAAAucGGcAAuGAATsT A18813 272 UUcAUUGCCGAUUUGUCGGTsTAD- 1089 11708 5886- L A18816 273 AGAucGAAAuuGuAcGAAGTsT A18817 274CUUCGuAcAAUUUCGAUCUTsT AD- 5904 11710  192-210 L A18818 275AAuccGcAAcuAcGcAAcuTsT A18819 276 AGUUGCGuAGUUGCGGAUUTsT AD- 11711 5395-L A18820 277 cAcGccAAuuAAcGucAucTsT A18821 278 GAUGACGUuAAUUGGCGUGTsTAD- 5413 11712  193-211 L A18822 279 AuccGcAAcuAcGcAAcuGTsT A18823 280cAGUUGCGuAGUUGCGGAUTsT AD- 11713  219-237 L A18824 281ccGAAAcAuAucuAccGuuTsT A18825 282 AACGGuAGAuAUGUUUCGGTsT AD- 11714 2840-L A18826 283 uuucuAccGGAAucuAGGATsT A18827 284 UCCuAGAUUCCGGuAGAAATsTAD- 2858 11715 4779- L A18828 285 AuuAAucGcGGAAcAAuuGTsT A18829 286cAAUUGUUCCGCGAUuAAUTsT AD- 4797 11716 5275- L A18830 287AuuucGAucGAucGAGAcATsT A18831 288 uGUCUCGAUCGAUCGAAAUTsT AD- 5293 117175391- L A18832 289 GGGAcAcGccAAuuAAcGuTsT A18833 290ACGUuAAUUGGCGUGUCCCTsT AD- 5409 11718  191-209 L A18834 291uAAuccGcAAcuAcGcAAcTsT A18835 292 GUUGCGuAGUUGCGGAUuATsT AD- 11719 1614-L A18836 293 AGuAcuAAAcGuGuAccGGTsT A18837 294 CCGGuAcACGUUuAGuACUTsTAD- 1632 11720 4588- L A18838 295 cAcAucGcucAuuGcGAAuTsT A18839 296AUUCGcAAUGAGCGAUGUGTsT AD- 4606 11721 4590- L A18840 297cAucGcucAuuGcGAAuAcTsT A18841 298 GuAUUCGcAAUGAGCGAUGTsT AD- 4608 117225884- L A18842 299 AcAGAucGAAAuuGuAcGATsT A18843 300UCGuAcAAUUUCGAUCUGUTsT AD- 5902 11723  161-179 L A18844 301AGcuuGcGGGuuAuAuucATsT A18845 302 UGAAuAuAACCCGcAAGCUTsT AD- 11724 778-796 L A18846 303 cuGccGAcGucuuGAuAAuTsT A18847 304AUuAUcAAGACGUCGGcAGTsT AD- 11725 5446- L A18848 305AGuAcuuAcGGcAAuuGAGTsT A18849 306 CUcAAUUGCCGuAAGuACUTsT AD- 5464 117266297- L A18850 307 AAccucGucGAuucAAAAATsT A18851 308uuuuuGAAUCGACGAGGuUTsT AD- 6315 11727 5269- L A18852 309AAcuAAAuuucGAucGAucTsT A18853 310 GAUCGAUCGAAAUUuAGUUTsT AD- 5287 117281778- L A18854 311 GccuuAuccGAcucGcAAuTsT A18855 312AUUGCGAGUCGGAuAAGGCTsT AD- 1796 11729 1780- L A18856 313cuuAuccGAcucGcAAuGuTsT A18857 314 AcAUUGCGAGUCGGAuAAGTsT AD- 1798 117303163- L A18858 315 GucGuuuuGcGGccGAuAuTsT A18859 316AuAUCGGCCGcAAAACGACTsT AD- 3181 11731 3164- L A18860 317ucGuuuuGcGGccGAuAucTsT A18861 318 GAuAUCGGCCGcAAAACGATsT AD- 3182 117325273- L A18862 319 AAAuuucGAucGAucGAGATsT A18863 320UCUCGAUCGAUCGAAAuuUTsT AD- 5291 11733 6295- L A18864 321AuAAccucGucGAuucAAATsT A18865 322 UUUGAAUCGACGAGGUuAUTsT AD- 6313 117341702- L A18866 323 uAcuAccAcAAuAucGGAATsT A18867 324UUCCGAuAUUGUGGuAGuATsT AD- 1720 11735 1781- L A18868 325uuAuccGAcucGcAAuGuuTsT A18869 326 AAcAUUGCGAGUCGGAuAATsT AD- 1799 117365270- L A18870 327 AcuAAAuuucGAucGAucGTsT A18871 328CGAUCGAUCGAAAUUuAGUTsT AD- 5288 11737 5276- L A18872 329uuucGAucGAucGAGAcAcTsT A18873 330 GuGUCUCGAUCGAUCGAAATsT AD- 5294 117385394- L A18874 331 AcAcGccAAuuAAcGucAuTsT A18875 332AUGACGUuAAUUGGCGUGUTsT AD- 5412 11739 6242- L A18876 333AAGuuAuAuccGccuuGGuTsT A18877 334 ACcAAGGCGGAuAuAACUUTsT AD- 6260 11740 182-200 L A18878 335 AuAcucccuuAAuccGcAATsT A18879 336UUGCGGAUuAAGGGAGuAUTsT AD- 11741  194-212 L A18880 337uccGcAAcuAcGcAAcuGuTsT A18881 338 AcAGUUGCGuAGUUGCGGATsT AD- 11742 575-593 L A18882 339 ucGAGGAAAcucuAGAucATsT A18883 340UGAUCuAGAGUUUCCUCGATsT AD- 11743 1565- L A18884 341uGcAGuAuucGAGccuAAuTsT A18885 342 AUuAGGCUCGAAuACUGcATsT AD- 1583 117441566- L A18886 343 GcAGuAuucGAGccuAAuGTsT A18887 344cAUuAGGCUCGAAuACUGCTsT AD- 1584 11745 1567- L A18888 345cAGuAuucGAGccuAAuGuTsT A18889 346 AcAUuAGGCUCGAAuACUGTsT AD- 1585 117462779- L A18890 347 cAuuGGcAcuAGcGGuAccTsT A18891 348GGuACCGCuAGUGCcAAUGTsT AD- 2797 11747 2838- L A18892 349uGuuucuAccGGAAucuAGTsT A18893 350 CuAGAUUCCGGuAGAAAcATsT AD- 2856 117482892- L A18894 351 AcuuAucuccGAAuGAuuGTsT A18895 352cAAUcAUUCGGAGAuAAGUTsT AD- 2910 11749 2981- L A18896 353AAAuccuAGcGGAuuAAAuTsT A18897 354 AUUuAAUCCGCuAGGAUUUTsT AD- 2999 117502982- L A18898 355 AAuccuAGcGGAuuAAAuGTsT A18899 356cAUUuAAUCCGCuAGGAUUTsT AD- 3000 11751 3038- L A18900 357GAuuGuAcGcAGGAccAucTsT A18901 358 GAUGGUCCUGCGuAcAAUCTsT AD- 3056 117523149- L A18902 359 AAcuccuGuuAuGAGucGuTsT A18903 360ACGACUcAuAAcAGGAGUUTsT AD- 3167 11753 3168- L A18904 361uuuGcGGccGAuAucuuuuTsT A18905 362 AAAAGAuAUCGGCCGcAAATsT AD- 3186 117543889- L A18906 363 GGuAcAAcGAucAAuAcAGTsT A18907 364CUGuAUUGAUCGUUGuACCTsT AD- 3907 11755 3922- L A18908 365uGGccAAucGuAuGAGuAATsT A18909 366 UuACUcAuACGAUUGGCcATsT AD- 3940 117564001- L A18910 367 GucuGcAcGcGAcAGcAAuTsT A18911 368AUUGCUGUCGCGUGcAGACTsT AD- 4019 11757 4584- L A18912 369cuAccAcAucGcucAuuGcTsT A18913 370 GcAAUGAGCGAUGUGGuAGTsT AD- 4602 117584593- L A18914 371 cGcucAuuGcGAAuAcuuATsT A18915 372uAAGuAUUCGcAAUGAGCGTsT AD- 4611 11759 4598- L A18916 373AuuGcGAAuAcuuAAGccATsT A18917 374 UGGCUuAAGuAUUCGcAAUTsT AD- 4616 117604601- L A18918 375 GcGAAuAcuuAAGccAAcATsT A18919 376UGUUGGCUuAAGuAUUCGCTsT AD- 4619 11761 4638- L A18920 377AuGucAcGGuuAAuGAGuATsT A18921 378 uACUcAUuAACCGUGAcAUTsT AD- 4656 117624778- L A18922 379 AAuuAAucGcGGAAcAAuuTsT A18923 380AAUUGUUCCGCGAUuAAUUTsT AD- 4796 11763 5274- L A18924 381AAuuucGAucGAucGAGAcTsT A18925 382 GUCUCGAUCGAUCGAAAuUTsT AD- 5292 117645392- L A18926 383 GGAcAcGccAAuuAAcGucTsT A18927 384GACGUuAAUUGGCGUGUCCTsT AD- 5410 11765 5649- L A18928 385AcGcuAGcuAcuGAGuccATsT A18929 386 UGGACUcAGuAGCuAGCGUTsT AD- 5667 117665833- L A18930 387 cuAAGcAAGucGAGGuuAuTsT A18931 388AuAACCUCGACUUGCUuAGTsT AD- 5851 11767 6243- L A18932 389AGuuAuAuccGccuuGGuuTsT A18933 390 AACcAAGGCGGAuAuAACUTsT AD- 6261 117686290- L A18934 391 cAGGuAuAAccucGucGAuTsT A18935 392AUCGACGAGGUuAuACCUGTsT AD- 6308 11769 6291- L A18936 393AGGuAuAAccucGucGAuuTsT A18937 394 AAUCGACGAGGUuAuACCUTsT AD- 6309 117701816- NP A18938 395 AcuAcGAGGAuucGGcuGATsT A18939 396UcAGCCGAAUCCUCGuAGUTsT AD- 1834 11771  875-893 NP A18940 397ucuAcccAAAcuuGucGuuTsT A18941 398 AACGAcAAGUUUGGGuAGATsT AD- 11772 1817-NP A18942 399 cuAcGAGGAuucGGcuGAATsT A18943 400 UUcAGCCGAAUCCUCGuAGTsTAD- 1835 11773 1812- NP A18944 401 ccuGAcuAcGAGGAuucGGTsT A18945 402CCGAAUCCUCGuAGUcAGGTsT AD- 1830 11774 1819- NP A18946 403AcGAGGAuucGGcuGAAGGTsT A18947 404 CCUUcAGCCGAAUCCUCGUTsT AD- 1837 117752140- NP A18948 405 AcGAGAGucucAcAucccuTsT A18949 406AGGGAuGuGAGACUCUCGUTsT AD- 2158 11776  730-748 VP35 A18950 407AAAuuucGGGcGAccuuAcTsT A18951 408 GuAAGGUCGCCCGAAAUUUTsT AD- 11777 735-753 VP35 A18952 409 ucGGGcGAccuuAcAuuucTsT A18953 410GAAAUGuAAGGUCGCCCGATsT AD- 11778  195-213 VP35 A18954 411uGAccGGcAAAAuAccGcuTsT A18955 412 AGCGGuAUUUUGCCGGUcATsT AD- 11779 198-216 VP35 A18956 413 ccGGcAAAAuAccGcuAAcTsT A18957 414GUuAGCGGuAUUUUGCCGGTsT AD- 11780  379-397 VP35 A18958 415AGcuGuGcGucGGcAAAccTsT A18959 416 GGUUUGCCGACGcAcAGCUTsT AD- 11781 646-664 VP35 A18960 417 AuuGAAAGAuccGAAcGGGTsT A18961 418CCCGUUCGGAUCUUUcAAUTsT AD- 11782  731-749 VP35 A18962 419AAuuucGGGcGAccuuAcATsT A18963 420 UGuAAGGUCGCCCGAAAUUTsT AD- 11783 732-750 VP35 A18964 421 AuuucGGGcGAccuuAcAuTsT A18965 422AUGuAAGGUCGCCCGAAAUTsT AD- 11784 1193- VP35 A18966 423GucuAuuGuGucAuAAGcuTsT A18967 424 AGCUuAUGAcAcAAuAGACTsT AD- 1211 11785 438-456 VP40 A18968 425 cucGcAucuuAuAcGAucATsT A18969 426UGAUCGuAuAAGAUGCGAGTsT AD- 11786 1301- VP40 A18970 427uGcAuAAGcGAuccAuAcuTsT A18971 428 AGuAUGGAUCGCUuAUGcATsT AD- 1319 117871191- VP40 A18972 429 AAuGuAcuAAucGGGucAATsT A18973 430UUGACCCGAUuAGuAcAUUTsT AD- 1209 11788  442-460 VP40 A18974 431cAucuuAuAcGAucAcccATsT A18975 432 UGGGUGAUCGuAuAAGAUGTsT AD- 11789 443-461 VP40 A18976 433 AucuuAuAcGAucAcccAuTsT A18977 434AUGGGUGAUCGuAuAAGAUTsT AD- 11790  478-496 VP40 A18978 435AcccccucGuuAGAGuGAATsT A18979 436 UUcACUCuAACGAGGGGGUTsT AD- 11791 834-852 VP40 A18980 437 AucGuGccAAuuGAuccAGTsT A18981 438CUGGAUcAAUUGGcACGAUTsT AD- 11792 1192- VP40 A18982 439AuGuAcuAAucGGGucAAGTsT A18983 440 CUUGACCCGAUuAGuAcAUTsT AD- 1210 117931194- VP40 A18984 441 GuAcuAAucGGGucAAGGATsT A18985 442UCCUUGACCCGAUuAGuACTsT AD- 1212 11794 1300- VP40 A18986 443AuGcAuAAGcGAuccAuAcTsT A18987 444 GuAUGGAUCGCUuAUGcAUTsT AD- 1318 11795 465-483 GP A18988 445 AcGGGAGcGAAuGcuuAccTsT A18989 446GGuAAGcAUUCGCUCCCGUTsT AD- 11796  358-376 VP30 A18990 447AGuuAGAGucccuAcGGuuTsT A18991 448 AACCGuAGGGACUCuAACUTsT AD- 11797 331-349 VP30 A18992 449 cuAccGuAGuAGucGAAGuTsT A18993 450ACUUCGACuACuACGGuAGTsT AD- 11798  250-268 VP30 A18994 451GAAuucAcGuGccGAccAGTsT A18995 452 CUGGUCGGcACGUGAAUUCTsT AD- 11799 1009-VP30 A18996 453 uGcccccccAAGcGuuAAuTsT A18997 454 AUuAACGCUUGGGGGGGcATsTAD- 1027 11800 1318- VP30 A18998 455 AGAGuGuuAGGAucGuuAuTsT A18999 456AuAACGAUCCuAAcACUCUTsT AD- 1336 11801  126-144 VP30 A19000 457AAucccGAGGcGGcAAuucTsT A19001 458 GAAuuGCCGCCUCGGGAuUTsT AD- 11802 354-372 VP30 A19002 459 cGcAAGuuAGAGucccuAcTsT A19003 460GuAGGGACUCuAACUUGCGTsT AD- 11803  553-571 VP30 A19004 461uGAuucAucGcuuAAuAuATsT A19005 462 uAuAUuAAGCGAUGAAUcATsT AD- 11804 583-601 VP30 A19006 463 AGAccuAAGAcuAGcAAAuTsT A19007 464AUUUGCuAGUCUuAGGUCUTsT AD- 11805  652-670 VP30 A19008 465AuuAcuAGucGAGAcuGcuTsT A19009 466 AGcAGUCUCGACuAGuAAUTsT AD- 11806 992-1010 VP30 A19010 467 ucAGGccuAcGcuuAcuuGTsT A19011 468cAAGuAAGCGuAGGCCUGATsT AD- 11807 1013- VP30 A19012 469cccccAAGcGuuAAuGAAGTsT A19013 470 CUUcAUuAACGCUUGGGGGTsT AD- 1031 11808 404-422 VP24 A19014 471 AuuAuAcGGGuccAuuAAuTsT A19015 472AUuAAUGGACCCGuAuAAUTsT AD- 11809  888-906 VP24 A19016 473cucAAcGAGuAAAGGAccATsT A19017 474 UGGUCCUUuACUCGUUGAGTsT AD- 11810 1247-VP24 A19018 475 uuGuAcGAuAGGGcuAAcATsT A19019 476 UGUuAGCCCuAUCGuAcAATsTAD- 1265 11811  536-554 VP24 A19020 477 GuuGuGuuuAGcGAccuAuTsT A19021478 AuAGGUCGCuAAAcAcAACTsT AD- 11812 1050- VP24 A19022 479GGAcuAAuAuGGGuuAucuTsT A19023 480 AGAuAACCcAuAUuAGUCCTsT AD- 1068 118131095- VP24 A19024 481 cuGcGAuGGAuAuAcGAcATsT A19025 482UGUCGuAuAUCcAUCGcAGTsT AD- 1113 11814  535-553 VP24 A19026 483AGuuGuGuuuAGcGAccuATsT A19027 484 uAGGUCGCuAAAcAcAACUTsT AD- 11815 196-214 VP24 A19028 485 uuGAAcuAGucuAcucGcATsT A19029 486UGCGAGuAGACuAGUUcAATsT AD- 11816  215-233 VP24 A19030 487GAAuccuAccGGGAAuAGATsT A19031 488 UCuAUUCCCGGuAGGAUUCTsT AD- 11817 403-421 VP24 A19032 489 uAuuAuAcGGGuccAuuAATsT A19033 490UuAAUGGACCCGuAuAAuATsT AD- 11818  406-424 VP24 A19034 491uAuAcGGGuccAuuAAuuuTsT A19035 492 AAAUuAAUGGACCCGuAuATsT AD- 11819 1140-VP24 A19036 493 uAcAuGAAucGAcAcuuAATsT A19037 494 UuAAGUGUCGAUUcAUGuATsTAD- 1158 11820 1243- VP24 A19038 495 AAAAuuGuAcGAuAGGGcuTsT A19039 496AGCCCuAUCGuAcAAUUUUTsT AD- 1261 11821 1249- VP24 A19040 497GuAcGAuAGGGcuAAcAuuTsT A19041 498 AAUGUuAGCCCuAUCGuACTsT AD- 1267 118221590- VP24 A19042 499 GAGcccAAAuuAAcAcGGuTsT A19043 500ACCGUGUuAAUUUGGGCUCTsT AD- 1608 11823 3688- L A19044 501cccGcuAuuAAGccGAGGuTsT A19045 502 ACCUCGGCUuAAuAGCGGGTsT AD- 3706 118243687- L A19046 503 GcccGcuAuuAAGccGAGGTsT A19047 504CCUCGGCUuAAuAGCGGGCTsT AD- 3705 11825 2956- L A19048 505AAuuGuAGcGcAAuuGAcuTsT A19049 506 AGUcAAUUGCGCuAcAAUUTsT AD- 2974 118262615- L A19050 507 AGcGAucAAucuccGAAAcTsT A19051 508GuuUCGGAGAuuGAUCGCUTsT AD- 2633 11827 2612- L A19052 509uuGAGcGAucAAucuccGATsT A19053 510 UCGGAGAUUGAUCGCUcAATsT AD- 2630 118284595- L A19054 511 uucGAAucuucAAAccGAcTsT A19055 512GUCGGuuuGAAGAuUCGAATsT AD- 4613 11829 2613- L A19056 513uGAGcGAucAAucuccGAATsT A19057 514 UUCGGAGAUUGAUCGCUcATsT AD- 2631 118302614- L A19058 515 GAGcGAucAAucuccGAAATsT A19059 516uuUCGGAGAuuGAUCGCUCTsT AD- 2632 11831 3941- L A19060 517cAAcGcGcuuGAuGGuAucTsT A19061 518 GAuACcAUcAAGCGCGUUGTsT AD- 3959 118323942- L A19062 519 AAcGcGcuuGAuGGuAucuTsT A19063 520AGAuACcAUcAAGCGCGUUTsT AD- 3960 11833 1680- L A19064 521AuAcGcccAAGAAcuuAGGTsT A19065 522 CCuAAGUUCUUGGGCGuAUTsT AD- 1698 118343686- L A19066 523 AGcccGcuAuuAAGccGAGTsT A19067 524CUCGGCUuAAuAGCGGGCUTsT AD- 3704 11835 4255- L A19068 525uuAucGAuuGAcAGuccuuTsT A19069 526 AAGGACUGUcAAUCGAuAATsT AD- 4273 118361374- L A19070 527 AGAccGAuGuuuAAcGccGTsT A19071 528CGGCGUuAAAcAUCGGUCUTsT AD- 1392 11837 5470- L A19072 529AccAuAuAuuGucGcuucATsT A19073 530 UGAAGCGAcAAuAuAUGGUTsT AD- 5488 118383872- L A19074 531 AuAuuGuGcAucGGuAuAATsT A19075 532UuAuACCGAUGcAcAAuAUTsT AD- 3890 11839 1384- L A19076 533uuAAcGccGGGAuuGAAuuTsT A19077 534 AAUUcAAUCCCGGCGUuAATsT AD- 1402 118404519- L A19078 535 uGcAcGAAAAAGAucGGAcTsT A19079 536GUCCGAUCUUUUUCGUGcATsT AD- 4537 11841 3682- L A19080 537GGucAGcccGcuAuuAAGcTsT A19081 538 GCUuAAuAGCGGGCUGACCTsT AD- 3700 118422954- L A19082 539 GGAAuuGuAGcGcAAuuGATsT A19083 540UcAAUUGCGCuAcAAUUCCTsT AD- 2972 11843 5467- L A19084 541AcuAccAuAuAuuGucGcuTsT A19085 542 AGCGAcAAuAuAUGGuAGUTsT AD- 5485 118441376- L A19086 543 AccGAuGuuuAAcGccGGGTsT A19087 544CCCGGCGUuAAAcAUCGGUTsT AD- 1394 11845 2448- L A19088 545uGAuGAGAcuuucGuAcAcTsT A19089 546 GUGuACGAAAGUCUcAUcATsT AD- 2466 118461023- L A19090 547 AcGAAAAGGGcGGuuuuuATsT A19091 548uAAAAACCGCCCUUUUCGUTsT AD- 1041 11847 1377- L A19092 549ccGAuGuuuAAcGccGGGATsT A19093 550 UCCCGGCGUuAAAcAUCGGTsT AD- 1395 118482619- L A19094 551 AucAAucuccGAAAcuAGATsT A19095 552UCuAGUUUCGGAGAUUGAUTsT AD- 2637 11849 5608- L A19096 553AAAuAcGGcGuuAAGAAGuTsT A19097 554 ACUUCUuAACGCCGuAUUUTsT AD- 5626 118505607- L A19098 555 AAAAuAcGGcGuuAAGAAGTsT A19099 556CUUCUuAACGCCGuAUUUUTsT AD- 5625 11851 6396- L A19100 557ucGAAcccAGAcuuAucAuTsT A19101 558 AUGAuAAGUCUGGGUUCGATsT AD- 6414 118524165- L A19102 559 AcAAccAcGcuAAAucuAGTsT A19103 560CuAGAUUuAGCGUGGUUGUTsT AD- 4183 11853 4250- L A19104 561GcAAcuuAucGAuuGAcAGTsT A19105 562 CUGUcAAUCGAuAAGUUGCTsT AD- 4268 118546434- L A19106 563 GAcGGAuAAcuAAAcuAGuTsT A19107 564ACuAGUUuAGUuAUCCGUCTsT AD- 6452 11855 2959- L A19108 565uGuAGcGcAAuuGAcuuuGTsT A19109 566 cAAAGUcAAUUGCGCuAcATsT AD- 2977 118566433- L A19110 567 GGAcGGAuAAcuAAAcuAGTsT A19111 568CuAGUUuAGUuAUCCGUCCTsT AD- 6451 11857   83-101 L A19112 569uGGcuAcccAAcAuAcAcATsT A19113 570 UGUGuAUGUUGGGuAGCcATsT AD- 11858 1382-L A19114 571 GuuuAAcGccGGGAuuGAATsT A19115 572 UUcAAUCCCGGCGUuAAACTsTAD- 1400 11859 1014- NP A19116 573 uuuccGuuuGAuGcGAAcATsT A19117 574UGUUCGcAUcAAACGGAAATsT AD- 1032 11860 1805- NP A19118 575GAAGcuAcGGcGAAuAccATsT A19119 576 UGGuAUUCGCCGuAGCUUCTsT AD- 1823 118611862- NP A19120 577 uGGuccuAuucGAucuAGATsT A19121 578UCuAGAUCGAAuAGGACcATsT AD- 1880 11862 1016- NP A19122 579uccGuuuGAuGcGAAcAAATsT A19123 580 UUUGUUCGcAUcAAACGGATsT AD- 1034 118632230- NP A19124 581 ccAccGGcucccGuAuAcATsT A19125 582UGuAuACGGGAGCCGGUGGTsT AD- 2248 11864 2233- NP A19126 583ccGGcucccGuAuAcAGAGTsT A19127 584 CUCUGuAuACGGGAGCCGGTsT AD- 2251 11865 959-977 NP A19136 585 AAGGAcuGAuAcAAuAuccTsT A19137 586GGAuAUUGuAUcAGUCCUUTsT AD- 11870 1017- NP A19138 587ccGuuuGAuGcGAAcAAAuTsT A19139 588 AUUUGUUCGcAUcAAACGGTsT AD- 1035 118712124- NP A19140 589 cccAcuGGAcGAuGccGAcTsT A19141 590GUCGGcAUCGUCcAGUGGGTsT AD- 2142 11872  745-763 NP A19142 591cGuGAuGGAGuGAAGcGccTsT A19143 592 GGCGCUUcACUCcAUcACGTsT AD- 11873 2229-NP A19144 593 cccAccGGcucccGuAuAcTsT A19145 594 GuAuACGGGAGCCGGUGGGTsTAD- 2247 11874 2119- NP A20118 595 GcAGAcccAcuGGAcGAuGTsT A20119 596cAUCGUCcAGUGGGUCUGCTsT AD- 2137 12462 1587- NP A20120 597AAAcGcuAuGGuAAcucuATsT A20121 598 uAGAGUuACcAuAGCGUUUTsT AD- 1605 124631300- NP A20122 599 uucGcccGAcuuuuGAAccTsT A20123 600GGUUcAAAAGUCGGGCGAATsT AD- 1318 12464 1808- NP A20124 601GcuAcGGcGAAuAccAGAGTsT A20125 602 CUCUGGuAUUCGCCGuAGCTsT AD- 1826 124651813- NP A20126 603 GGcGAAuAccAGAGuuAcuTsT A20127 604AGuAACUCUGGuAUUCGCCTsT AD- 1831 12466  532-550 VP35 A19146 605GGuGAuGAcAAccGGucGGTsT A19147 606 CCGACCGGUUGUcAUcACCTsT AD- 11875 417-435 VP35 A19148 607 uAGAAcAAcGcAuuAcGAGTsT A19149 608CUCGuAAUGCGUUGUUCuATsT AD- 11876  741-759 VP35 A19152 609GGAAAccuGAcAuuucGGcTsT A19153 610 GCCGAAAUGUcAGGUUUCCTsT AD- 11878 1049-VP35 A19154 611 cccAAGAuuGAucGAGGuuTsT A19155 612 AACCUCGAUcAAUCUUGGGTsTAD- 1067 11879  206-224 VP35 A19160 613 AGAAuuccuGuAAGcGAcATsT A19161614 UGUCGCUuAcAGGAAUUCUTsT AD- 11882  246-264 VP35 A19162 615AuccAGGAuuAuGcuAcGcTsT A19163 616 GCGuAGcAuAAUCCUGGAUTsT AD- 11883 247-265 VP35 A19164 617 uccAGGAuuAuGcuAcGcATsT A19165 618UGCGuAGcAuAAUCCUGGATsT AD- 11884  287-305 VP35 A19166 619ccAAAcccGAAGAcGcGcATsT A19167 620 uGCGCGUCuUCGGGuuuGGTsT AD- 11885 314-332 VP35 A19168 621 AcccAAAcGGAcccAAuuuTsT A19169 622AAAuuGGGUCCGuuuGGGUTsT AD- 11886  319-337 VP35 A19170 623AAcGGAcccAAuuuGcAAuTsT A19171 624 AUUGcAAAUUGGGUCCGUUTsT AD- 11887 414-432 VP35 A19172 625 cAuuAGAAcAAcGcAuuAcTsT A19173 626GuAAUGCGUUGUUCuAAUGTsT AD- 11888  415-433 VP35 A19174 627AuuAGAAcAAcGcAuuAcGTsT A19175 628 CGuAAUGCGUUGUUCuAAUTsT AD- 11889 439-457 VP35 A19176 629 uGAGAAuGGucuAAAGccATsT A19177 630UGGCUUuAGACcAUUCUcATsT AD- 11890  576-594 VP35 A19178 631AGGcuuAuuGGGccGAAcATsT A19179 632 UGUUCGGCCcAAuAAGCCUTsT AD- 11891 413-431 VP35 A20128 633 ucAuuAGAAcAAcGcAuuATsT A20129 634uAAUGCGUUGUUCuAAUGATsT AD- 12467  583-601 VP35 A20130 635uuGGGccGAAcAuGGucAATsT A20131 636 UUGACcAUGUUCGGCCcAATsT AD- 12468 983-1001 VP35 A20132 637 cAcAuccGcucucGAGGuGTsT A20133 638cACCUCGAGAGCGGAUGUGTsT AD- 12469  318-336 VP35 A20134 639AAAcGGAcccAAuuuGcAATsT A20135 640 UUGcAAAUUGGGUCCGUUUTsT AD- 12470 420-438 VP35 A20136 641 AAcAAcGcAuuAcGAGucuTsT A20137 642AGACUCGuAAUGCGUUGUUTsT AD- 12471  419-437 VP35 A20138 643GAAcAAcGcAuuAcGAGucTsT A20139 644 GACUCGuAAUGCGUUGUUCTsT AD- 12472 134-152 VP35 A20140 645 GccAcGAcucAAAAcGAcATsT A20141 646UGUCGUUUUGAGUCGUGGCTsT AD- 12473  893-911 VP40 A19180 647ccAcAAGcuGAccGGuAAGTsT A19181 648 CUuACCGGUcAGCUUGUGGTsT AD- 11892 892-910 VP40 A19182 649 uccAcAAGcuGAccGGuAATsT A19183 650UuACCGGUcAGCUUGUGGATsT AD- 11893  325-343 VP40 A19188 651uGAAuGucAuAucGGGcccTsT A19189 652 GGGCCCGAuAUGAcAUUcATsT AD- 11896 450-468 VP40 A19190 653 AcuAucAcccAuuucGGcATsT A19191 654UGCCGAAAUGGGUGAuAGUTsT AD- 11897  662-680 VP40 A19194 655GAccGAuGAcAcuccAAcATsT A19195 656 UGUUGGAGUGUcAUCGGUCTsT AD- 11899 200-218 VP40 A19198 657 GAcAccGGAGucAGucAAuTsT A19199 658AuuGACuGACUCCGGuGUCTsT AD- 11901  203-221 VP40 A19200 659AccGGAGucAGucAAuGGGTsT A19201 660 CCcAUUGACUGACUCCGGUTsT AD- 11902 204-222 VP40 A19202 661 ccGGAGucAGucAAuGGGGTsT A19203 662CCCcAUUGACUGACUCCGGTsT AD- 11903  225-243 VP40 A19204 663AcuccAucGAAuccAcucATsT A19205 664 uGAGuGGAuUCGAuGGAGUTsT AD- 11904 386-404 VP40 A19206 665 uGucGcuGAucAAAAGAccTsT A19207 666GGUCUUUUGAUcAGCGAcATsT AD- 11905  584-602 VP40 A19208 667AGuccAAcuAccccAGuAuTsT A19209 668 AuACUGGGGuAGUUGGACUTsT AD- 11906 631-649 VP40 A19210 669 uGAucAcccAAccAcuGccTsT A19211 670GGcAGUGGUUGGGUGAUcATsT AD- 11907  660-678 VP40 A19212 671uGGAccGAuGAcAcuccAATsT A19213 672 UUGGAGUGUcAUCGGUCcATsT AD- 11908 663-681 VP40 A19214 673 AccGAuGAcAcuccAAcAGTsT A19215 674CUGUUGGAGUGUcAUCGGUTsT AD- 11909  929-947 VP40 A19218 675uGGAcAAccAAucAucccuTsT A19219 676 AGGGAUGAUUGGUUGUCcATsT AD- 11911 1019-VP40 A19220 677 uuGuGAcAcGuGucAuucuTsT A19221 678 AGAAUGAcACGUGUcAcAATsTAD- 1037 11912  243-261 VP40 A19224 679 AGGccAAuuGccGAuGAcATsT A19225680 UGUcAUCGGcAAUUGGCCUTsT AD- 11914  140-158 VP40 A20142 681AuAcccuGucAGGucAAAuTsT A20143 682 AUUUGACCUGAcAGGGuAUTsT AD- 12474 141-159 VP40 A20144 683 uAcccuGucAGGucAAAuuTsT A20145 684AAUUUGACCUGAcAGGGuATsT AD- 12475  378-396 VP40 A20146 685ccucuAGGuGucGcuGAucTsT A20147 686 GAUcAGCGAcACCuAGAGGTsT AD- 12476 427-445 VP40 A20148 687 ccGccAucAuGcuuGcuucTsT A20149 688GAAGcAAGcAUGAUGGCGGTsT AD- 12477  898-916 VP40 A20150 689AGcuGAccGGuAAGAAGGuTsT A20151 690 ACCUUCUuACCGGUcAGCUTsT AD- 12478 199-217 VP40 A20152 691 uGAcAccGGAGucAGucAATsT A20153 692UUGACUGACUCCGGUGUcATsT AD- 12479  568-586 VP40 A20154 693AGuucGuucuuccGccAGuTsT A20155 694 ACUGGCGGAAGAACGAACUTsT AD- 12480 569-587 VP40 A20156 695 GuucGuucuuccGccAGucTsT A20157 696GACUGGCGGAAGAACGAACTsT AD- 12481 1728- GP A19232 697ccuGGAuAccAuAuuucGGTsT A19233 698 CCGAAAuAUGGuAUCcAGGTsT AD- 1746 119181729- GP A19234 699 cuGGAuAccAuAuuucGGGTsT A19235 700CCCGAAAuAUGGuAUCcAGTsT AD- 1747 11919 1818- GP A19246 701AGcuGGccAAcGAGAcGAcTsT A19247 702 GUCGUCUCGUUGGCcAGCUTsT AD- 1836 119251821- GP A19248 703 uGGccAAcGAGAcGAcucATsT A19249 704UGAGUCGUCUCGUUGGCcATsT AD- 1839 11926 1732- GP A19250 705GAuAccAuAuuucGGGccATsT A19251 706 UGGCCCGAAAuAUGGuAUCTsT AD- 1750 119271956- GP A20158 707 cGGAcuGcuGuAucGAAccTsT A20159 708GGUUCGAuAcAGcAGUCCGTsT AD- 1974 12482 2107- GP A20160 709uGGAGuuAcAGGcGuuAuATsT A20161 710 uAuAACGCCUGuAACUCcATsT AD- 2125 124832124- GP A20162 711 uAAuuGcAGuuAucGcuuuTsT A20163 712AAAGCGAuAACUGcAAUuATsT AD- 2142 12484 2109- GP A20164 713GAGuuAcAGGcGuuAuAAuTsT A20165 714 AUuAuAACGCCUGuAACUCTsT AD- 2127 124851958- GP A20166 715 GAcuGcuGuAucGAAccAcTsT A20167 716GUGGUUCGAuAcAGcAGUCTsT AD- 1976 12486 1890- GP A20168 717uccucAAccGuAAGGcAAuTsT A20169 718 AUUGCCUuACGGUUGAGGATsT AD- 1908 124871891- GP A20170 719 ccucAAccGuAAGGcAAuuTsT A20171 720AAUUGCCUuACGGUUGAGGTsT AD- 1909 12488 1307- GP A20172 721AAuAcAcccGuGuAuAAAcTsT A20173 722 GUUuAuAcACGGGUGuAUUTsT AD- 1325 124891823- GP A20174 723 GccAAcGAGAcGAcucAAGTsT A20175 724CUUGAGUCGUCUCGUUGGCTsT AD- 1841 12490 2110- GP A20176 725AGuuAcAGGcGuuAuAAuuTsT A20177 726 AAUuAuAACGCCUGuAACUTsT AD- 2128 124911308- GP A20178 727 AuAcAcccGuGuAuAAAcuTsT A20179 728AGUUuAuAcACGGGUGuAUTsT AD- 1326 12492 2113- GP A20180 729uAcAGGcGuuAuAAuuGcATsT A20181 730 UGcAAUuAuAACGCCUGuATsT AD- 2131 124931654- GP A20182 731 cAAuGcucAAcccAAAuGcTsT A20183 732GcAUUUGGGUUGAGcAUUGTsT AD- 1672 12494 1824- GP A20184 733ccAAcGAGAcGAcucAAGcTsT A20185 734 GCUUGAGUCGUCUCGUUGGTsT AD- 1842 124951313- GP A20186 735 cccGuGuAuAAAcuuGAcATsT A20187 736UGUcAAGUUuAuAcACGGGTsT AD- 1331 12496 1873- GP A20188 737GcuAcGcAccuuuucAAucTsT A20189 738 GAUUGAAAAGGUGCGuAGCTsT AD- 1891 124971953- GP A20190 739 GAccGGAcuGcuGuAucGATsT A20191 740UCGAuAcAGcAGUCCGGUCTsT AD- 1971 12498 1964- GP A20192 741uGuAucGAAccAcAuGAuuTsT A20193 742 AAUcAUGUGGUUCGAuAcATsT AD- 1982 12499 329-347 VP30 A20194 743 AGGuGAGuAccGucAAucATsT A20195 744UGAUUGACGGuACUcACCUTsT AD- 12500  426-444 VP30 A20196 745AAAGAcAuAuGuccGAccuTsT A20197 746 AGGUCGGAcAuAUGUCUUUTsT AD- 12501 842-860 VP30 A20198 747 ucucGAAGuAuAucAAcGATsT A20199 748UCGUUGAuAuACUUCGAGATsT AD- 12502  909-927 VP30 A20200 749uGGGAccGAcAAucccuAATsT A20201 750 UuAGGGAUUGUCGGUCCcATsT AD- 12503 523-541 VP30 A20202 751 uccuAcuAAucGcccGuAATsT A20203 752UuACGGGCGAUuAGuAGGATsT AD- 12504  429-447 VP30 A20204 753GAcAuAuGuccGAccuuGATsT A20205 754 UcAAGGUCGGAcAuAUGUCTsT AD- 12505 521-539 VP30 A20206 755 AcuccuAcuAAucGcccGuTsT A20207 756ACGGGCGAUuAGuAGGAGUTsT AD- 12506  903-921 VP30 A20208 757cAAcAAuGGGAccGAcAAuTsT A20209 758 AUUGUCGGUCCcAUUGUUGTsT AD- 12507 355-373 VP30 A20210 759 ccucAcAAGuGcGcGuuccTsT A20211 760GGAACGCGcACUUGUGAGGTsT AD- 12508  337-355 VP30 A20212 761AccGucAAucAAGGAGcGcTsT A20213 762 GCGCUCCUUGAUUGACGGUTsT AD- 12509 908-926 VP24 A19262 763 cuGucGuuGAuucGAuccATsT A19263 764UGGAUCGAAUcAACGAcAGTsT AD- 11933  522-540 VP24 A19272 765uuGucuuAAGcGAccucuGTsT A19273 766 cAGAGGUCGCUuAAGAcAATsT AD- 11938 790-808 VP24 A19274 767 uuuGAuuGAAcccuuAGcATsT A19275 768UGCuAAGGGUUcAAUcAAATsT AD- 11939  863-881 VP24 A20214 769AAcAuGcGAAcAcAAcGuGTsT A20215 770 cACGUUGUGUUCGcAUGUUTsT AD- 12510 1102-VP24 A20216 771 uGGGccGGcGAAAuuuuccTsT A20217 772 GGAAAAUUUCGCCGGCCcATsTAD- 1120 12511  912-930 VP24 A20218 773 cGuuGAuucGAuccAAuAuTsT A20219774 AuAUUGGAUCGAAUcAACGTsT AD- 12512  954-972 VP24 A20220 775AuGcucuAcAuGucGuGAATsT A20221 776 UUcACGAcAUGuAGAGcAUTsT AD- 12513 475-493 VP24 A20222 777 GGGAcGAuAcAAucuAAuATsT A20223 778uAUuAGAUUGuAUCGUCCCTsT AD- 12514 1069- VP24 A20224 779AcccGAcAAAucGGcAAuGTsT A20225 780 cAUUGCCGAUUUGUCGGGUTsT AD- 1087 12515 486-504 VP24 A20226 781 AucuAAuAucGcccAAAAATsT A20227 782UUUUUGGGCGAuAUuAGAUTsT AD- 12516  525-543 VP24 A20228 783ucuuAAGcGAccucuGuAATsT A20229 784 UuAcAGAGGUCGCUuAAGATsT AD- 12517 867-885 VP24 A20230 785 uGcGAAcAcAAcGuGucAATsT A20231 786UUGAcACGUUGUGUUCGcATsT AD- 12518 1028- VP24 A20232 787AuAAcucGAAcuAAcAuGGTsT A20233 788 CcAUGUuAGUUCGAGUuAUTsT AD- 1046 12519 471-489 VP24 A20234 789 cuAcGGGAcGAuAcAAucuTsT A20235 790AGAUUGuAUCGUCCCGuAGTsT AD- 12520 1029- VP24 A20236 791uAAcucGAAcuAAcAuGGGTsT A20237 792 CCcAUGUuAGUUCGAGUuATsT AD- 1047 125211948- L A20238 793 cAGuuAGAGGGAGuAGcuuTsT A20239 794AAGCuACUCCCUCuAACUGTsT AD- 1966 12522 2003- L A20240 795AuAuGAGuuuAcAGcAccuTsT A20241 796 AGGUGCUGuAAACUcAuAUTsT AD- 2021 125232005- L A20242 797 AuGAGuuuAcAGcAccuuuTsT A20243 798AAAGGUGCUGuAAACUcAUTsT AD- 2023 12524 2070- L A20244 799uGGAuGcAuuAuAcAAuccTsT A20245 800 GGAUUGuAuAAUGcAUCcATsT AD- 2088 125251959- NP A19278 801 AccuuGGAcGGAGcGAAAATsT A19279 802UUUUCGCUCCGUCcAAGGUTsT AD- 1977 11941 1687- NP A19280 803cAuuucccGGGccGAucuATsT A19281 804 uAGAUCGGCCCGGGAAAUGTsT AD- 1705 119421775- NP A19282 805 uGuuGuuGAcccGuAuGAuTsT A19283 806AUcAuACGGGUcAAcAAcATsT AD- 1793 11943  384-402 NP A19284 807GuuuAccuGAGAGccuAcATsT A19285 808 UGuAGGCUCUcAGGuAAACTsT AD- 11944 400-418 NP A19286 809 AcAAcAuGGAuAAAcGGGuTsT A19287 810ACCCGUUuAUCcAUGUUGUTsT AD- 11945 1773- NP A19288 811GGuGuuGuuGAcccGuAuGTsT A19289 812 cAuACGGGUcAAcAAcACCTsT AD- 1791 119461964- NP A19290 813 GGAcGGAGcGAAAAAGGuGTsT A19291 814cACCUUUUUCGCUCCGUCCTsT AD- 1982 11947  411-429 NP A19292 815AAAcGGGuGAGAGGuucAuTsT A19293 816 AUGAACCUCUcACCCGUUUTsT AD- 11948 1815-NP A19294 817 GAcuAcGAGGAuucGGcuGTsT A19295 818 cAGCCGAAUCCUCGuAGUCTsTAD- 1833 11949  407-425 NP A19296 819 GGAuAAAcGGGuGAGAGGuTsT A19297 820ACCUCUcACCCGUUuAUCCTsT AD- 11950 2405- NP A19298 821uuAucAccuAAuGAGuGAuTsT A19299 822 AUcACUcAUuAGGUGAuAATsT AD- 2423 11951 409-427 NP A19300 823 AuAAAcGGGuGAGAGGuucTsT A19301 824GAACCUCUcACCCGUUuAUTsT AD- 11952 1811- NP A19302 825uccuGAcuAcGAGGAuucGTsT A19303 826 CGAAUCCUCGuAGUcAGGATsT AD- 1829 11953 408-426 NP A19304 827 GAuAAAcGGGuGAGAGGuuTsT A19305 828AACCUCUcACCCGUUuAUCTsT AD- 11954 1958- NP A19306 829GAccuuGGAcGGAGcGAAATsT A19307 830 UUUCGCUCCGUCcAAGGUCTsT AD- 1976 119551973- NP A19308 831 GAAAAAGGuGccGGAGuuGTsT A19309 832cAACUCCGGcACCUUUUUCTsT AD- 1991 11956 1810- NP A19310 833AuccuGAcuAcGAGGAuucTsT A19311 834 GAAUCCUCGuAGUcAGGAUTsT AD- 1828 119571953- NP A19312 835 GAuccGAccuuGGAcGGAGTsT A19313 836CUCCGUCcAAGGUCGGAUCTsT AD- 1971 11958 1692- NP A19314 837cccGGGccGAucuAuGAuGTsT A19315 838 cAUcAuAGAUCGGCCCGGGTsT AD- 1710 11959 197-215 VP35 A19316 839 AccGGcAAAAuAccGcuAATsT A19317 840UuAGCGGuAUUUUGCCGGUTsT AD- 11960  196-214 VP35 A19318 841GAccGGcAAAAuAccGcuATsT A19319 842 uAGCGGuAUUUUGCCGGUCTsT AD- 11961 409-427 VP35 A19320 843 AucAcuAGAAGGucGAGuATsT A19321 844uACUCGACCUUCuAGUGAUTsT AD- 11962  476-494 VP35 A19322 845AuAucAucccuGAAucGcATsT A19323 846 UGCGAUUcAGGGAUGAuAUTsT AD- 11963 611-629 VP35 A19324 847 ccAucAuuGuAcGAGGAuGTsT A19325 848cAUCCUCGuAcAAUGAUGGTsT AD- 11964  645-663 VP35 A19326 849AAuuGAAAGAuccGAAcGGTsT A19327 850 CCGUUCGGAUCUUUcAAUUTsT AD- 11965 726-744 VP35 A19328 851 AGGAAAAuuucGGGcGAccTsT A19329 852GGUCGCCCGAAAuuuUCCUTsT AD- 11966 1130- VP35 A19330 853GuAAGcucAuuuuGcGAuGTsT A19331 854 cAUCGcAAAAUGAGCUuACTsT AD- 1148 11967 729-747 VP35 A19332 855 AAAAuuucGGGcGAccuuATsT A19333 856uAAGGUCGCCCGAAAUUUUTsT AD- 11968  606-624 VP35 A19334 857cAGGcccAucAuuGuAcGATsT A19335 858 UCGuAcAAUGAUGGGCCUGTsT AD- 11969 256-274 VP35 A19336 859 cccGAuAAccAuuAuuAGuTsT A19337 860ACuAAuAAUGGUuAUCGGGTsT AD- 11970  478-496 VP35 A19338 861AucAucccuGAAucGcAGcTsT A19339 862 GCUGCGAUUcAGGGAUGAUTsT AD- 11971 724-742 VP35 A19340 863 uGAGGAAAAuuucGGGcGATsT A19341 864UCGCCCGAAAUUUUCCUcATsT AD- 11972  644-662 VP35 A19342 865AAAuuGAAAGAuccGAAcGTsT A19343 866 CGUUCGGAUCUUUcAAUUUTsT AD- 11973 1239-VP35 A19344 867 AuccuAAucAAuuGAuAAuTsT A19345 868 AUuAUcAAUUGAUuAGGAUTsTAD- 1257 11974 1052- VP35 A19346 869 AucGAuAGAGGuuGGGucuTsT A19347 870AGACCcAACCUCuAUCGAUTsT AD- 1070 11975  429-447 VP40 A19348 871GcAAuuAuGcucGcAucuuTsT A19349 872 AAGAUGCGAGcAuAAUUGCTsT AD- 11976 1189-VP40 A19350 873 AAAAuGuAcuAAucGGGucTsT A19351 874 GACCCGAUuAGuAcAUUUUTsTAD- 1207 11977 1190- VP40 A19352 875 AAAuGuAcuAAucGGGucATsT A19353 876UGACCCGAUuAGuAcAUUUTsT AD- 1208 11978  373-391 VP40 A19354 877GGuuGccAcucGGAAuuGcTsT A19355 878 GcAAUUCCGAGUGGcAACCTsT AD- 11979 439-457 VP40 A19356 879 ucGcAucuuAuAcGAucAcTsT A19357 880GUGAUCGuAuAAGAUGCGATsT AD- 11980  441-459 VP40 A19358 881GcAucuuAuAcGAucAcccTsT A19359 882 GGGUGAUCGuAuAAGAUGCTsT AD- 11981 1121-VP40 A19360 883 AuAGcAAcucAAucGAcuuTsT A19361 884 AAGUCGAUUGAGUUGCuAUTsTAD- 1139 11982 1127- VP40 A19362 885 AcucAAucGAcuuuuAGGATsT A19363 886UCCuAAAAGUCGAUUGAGUTsT AD- 1145 11983 1193- VP40 A19364 887uGuAcuAAucGGGucAAGGTsT A19365 888 CCUUGACCCGAUuAGuAcATsT AD- 1211 119841298- VP40 A19366 889 AcAuGcAuAAGcGAuccAuTsT A19367 890AUGGAUCGCUuAUGcAUGUTsT AD- 1316 11985 1307- VP40 A19368 891AGcGAuccAuAcuucGcccTsT A19369 892 GGGCGAAGuAUGGAUCGCUTsT AD- 1325 11986 361-379 VP40 A19370 893 AAAucccuAuuuGGuuGccTsT A19371 894GGcAACcAAAuAGGGAUUUTsT AD- 11987  437-455 VP40 A19372 895GcucGcAucuuAuAcGAucTsT A19373 896 GAUCGuAuAAGAUGCGAGCTsT AD- 11988 857-875 VP40 A19374 897 GAGuAucAuuGGGAucGAGTsT A19375 898CUCGAUCCcAAUGAuACUCTsT AD- 11989  484-502 VP40 A19376 899ucGuuAGAGuGAAucGAcuTsT A19377 900 AGUCGAUUcACUCuAACGATsT AD- 11990 1845-GP A19378 901 GGAGcuGcGGAcAuAuAccTsT A19379 902 GGuAuAUGUCCGcAGCUCCTsTAD- 1863 11991  254-272 GP A19380 903 GAGAuuGAccAGcuAGucuTsT A19381 904AGACuAGCUGGUcAAUCUCTsT AD- 11992  461-479 GP A19382 905ccGGAcGGGAGcGAAuGcuTsT A19383 906 AGcAUUCGCUCCCGUCCGGTsT AD- 11993 466-484 GP A19384 907 cGGGAGcGAAuGcuuAcccTsT A19385 908GGGuAAGcAUUCGCUCCCGTsT AD- 11994  933-951 GP A19386 909uAAuuuGGAcAcuAGAuGcTsT A19387 910 GcAUCuAGUGUCcAAAUuATsT AD- 11995 1045-GP A19388 911 uuAucGcucAAcGAGAcAGTsT A19389 912 CUGUCUCGUUGAGCGAuAATsTAD- 1063 11996 1100- GP A19390 913 GAAGAAucuccGAccGGGcTsT A19391 914GCCCGGUCGGAGAuUCuUCTsT AD- 1118 11997 1102- GP A19392 915AGAAucuccGAccGGGccATsT A19393 916 UGGCCCGGUCGGAGAUUCUTsT AD- 1120 119981191- GP A19394 917 AAcAAcAuuGccGucucAGTsT A19395 918CUGAGACGGcAAUGUUGUUTsT AD- 1209 11999 1203- GP A19396 919GucucAGAAuucGAcAGAATsT A19397 920 uUCuGUCGAAuUCuGAGACTsT AD- 1221 120001844- GP A19398 921 cGGAGcuGcGGAcAuAuAcTsT A19399 922GuAuAUGUCCGcAGCUCCGTsT AD- 1862 12001  255-273 GP A19400 923AGAuuGAccAGcuAGucuGTsT A19401 924 cAGACuAGCUGGUcAAUCUTsT AD- 12002 1212-GP A19402 925 uucGAcAGAAGGucGAAGATsT A19403 926 UCUUCGACCUUCUGUCGAATsTAD- 1230 12003 1706- GP A19404 927 GGAucccGuAcuuuGGAccTsT A19405 928GGUCcAAAGuACGGGAUCCTsT AD- 1724 12004  125-143 GP A19406 929cuuAGccuAcuccAAuuGcTsT A19407 930 GcAAUUGGAGuAGGCuAAGTsT AD- 12005 264-282 GP A19408 931 AGcuAGucuGcAAGGAucATsT A19409 932UGAUCCUUGcAGACuAGCUTsT AD- 12006  332-350 GP A19410 933AGcGGAGuAucuAcuGAuATsT A19411 934 uAUcAGuAGAuACUCCGCUTsT AD- 12007 464-482 GP A19412 935 GAcGGGAGcGAAuGcuuAcTsT A19413 936GuAAGcAUUCGCUCCCGUCTsT AD- 12008 1210- GP A19414 937AAuucGAcAGAAGGucGAATsT A19415 938 uUCGACCuUCuGUCGAAUUTsT AD- 1228 120091213- GP A19416 939 ucGAcAGAAGGucGAAGAGTsT A19417 940CUCuUCGACCuUCuGUCGATsT AD- 1231 12010 1850- GP A19418 941uGcGGAcAuAuAccAuAcuTsT A19419 942 AGuAUGGuAuAUGUCCGcATsT AD- 1868 12011 124-142 GP A19420 943 ucuuAGccuAcuccAAuuGTsT A19421 944cAAUUGGAGuAGGCuAAGATsT AD- 12012 1044- GP A19422 945uuuAucGcucAAcGAGAcATsT A19423 946 UGUCUCGUUGAGCGAuAAATsT AD- 1062 12013 265-283 GP A19424 947 GcuAGucuGcAAGGAucAuTsT A19425 948AUGAUCCUUGcAGACuAGCTsT AD- 12014  361-379 VP30 A19426 949uAGAGucccuAcGGuuuucTsT A19427 950 GAAAACCGuAGGGACUCuATsT AD- 12015 324-342 VP30 A19428 951 cAAcAGAcuAccGuAGuAGTsT A19429 952CuACuACGGuAGUCUGUUGTsT AD- 12016  994-1012 VP30 A14390 953AGGccuAcGcuuAcuuGccTsT A19431 954 GGcAAGuAAGCGuAGGCCUTsT AD- 12017 248-266 VP30 A19432 955 AGGAAuucAcGuGccGAccTsT A19433 956GGUCGGcACGUGAAUUCCUTsT AD- 12018  491-509 VP30 A19434 957cuuGAAAGccuAAccGAccTsT A19435 958 GGUCGGUuAGGCUUUcAAGTsT AD- 12019 322-340 VP30 A19436 959 AAcAAcAGAcuAccGuAGuTsT A19437 960ACuACGGuAGUCUGUUGUUTsT AD- 12020  323-341 VP30 A19438 961AcAAcAGAcuAccGuAGuATsT A19439 962 uACuACGGuAGUCUGUUGUTsT AD- 12021 517-535 VP30 A19440 963 ccuAcuucuuAuAGcAcGGTsT A19441 964CCGUGCuAuAAGAAGuAGGTsT AD- 12022  295-313 VP30 A19442 965GAcAAGAuccAuuucccGGTsT A19443 966 CCGGGAAAuGGAUCuuGUCTsT AD- 12023 229-247 VP30 A19444 967 ucGuGAGcGcGGGAGAucATsT A19445 968UGAUCUCCCGCGCUcACGATsT AD- 12024  251-269 VP30 A19446 969AAuucAcGuGccGAccAGcTsT A19447 970 GCUGGUCGGcACGUGAAUUTsT AD- 12025 340-358 VP30 A19448 971 uAGucGAAGuAcuucGcAATsT A19449 972UUGCGAAGuACUUCGACuATsT AD- 12026 1350- VP30 A19450 973ucccuAGAAGcGuuGAAucTsT A19451 974 GAUUcAACGCUUCuAGGGATsT AD- 1368 120271057- VP24 A19452 975 uAuGGGuuAucuuGucGAGTsT A19453 976CUCGAcAAGAuAACCcAuATsT AD- 1075 12028  878-896 VP24 A19454 977AAcAuGAGAAcucAAcGAGTsT A19455 978 CUCGUUGAGUUCUcAUGUUTsT AD- 12029 1056-VP24 A19456 979 AuAuGGGuuAucuuGucGATsT A19457 980 UCGAcAAGAuAACCcAuAUTsTAD- 1074 12030 1137- VP24 A19458 981 uAcuAcAuGAAucGAcAcuTsT A19459 982AGUGUCGAUUcAUGuAGuATsT AD- 1155 12031 1099- VP24 A19460 983GAuGGAuAuAcGAcAcccuTsT A19461 984 AGGGUGUCGuAuAUCcAUCTsT AD- 1117 120321591- VP24 A19462 985 AGcccAAAuuAAcAcGGuATsT A19463 986uACCGUGUuAAUUUGGGCUTsT AD- 1609 12033 1094- VP24 A19464 987ucuGcGAuGGAuAuAcGAcTsT A19465 988 GUCGuAuAUCcAUCGcAGATsT AD- 1112 120341135- VP24 A19466 989 cuuAcuAcAuGAAucGAcATsT A19467 990UGUCGAUUcAUGuAGuAAGTsT AD- 1153 12035  152-170 VP24 A19468 991cuAGGcuAGGGuuuAuAGuTsT A19469 992 ACuAuAAACCCuAGCCuAGTsT AD- 12036 624-642 VP24 A19470 993 AccAAAAGGGuAuuAcccuTsT A19471 994AGGGuAAuACCCUUUUGGUTsT AD- 12037

TABLE 3 Ebola Ebola Ebola Zaire GFP assay Zaire plaque Ebola Sudan IFassay Sudan plaque duplex % reduction % reduction vs % reduction %reduction vs name 100 nM 10 nM 1 nM no siRNA 100 nM 10 nM 1 nM no siRNAAD-11542 90.7 80.1 55.2 55.24 −5.3 −1.5 −11.8 −103.39 AD-11543 62.7 55.947.8 55.83 −11.9 10.2 −9.0 −7.12 AD-11544 63.5 32.8 41.0 42.06 22.5 34.661.1 −16.95 AD-11545 73.0 71.8 61.4 47.23 61.5 71.9 71.8 39.41 AD-1154694.5 88.3 72.5 86.12 −8.8 −73.8 5.5 55.93 AD-11547 79.8 62.8 48.6 36.44−20.7 15.8 −4.0 18.64 AD-11548 −15.4 −13.0 −15.6 19.3 29.2 49.3 AD-115493.8 13.8 16.1 44.12 63.7 51.5 90.4 94.38 AD-11550 17.5 −1.2 −10.5 −41.1−39.8 −29.4 AD-11551 −14.2 −20.4 −21.3 −32.6 −23.9 −30.6 AD-11552 −14.7−11.9 −6.5 −22.7 −9.5 11.9 AD-11553 4.7 7.4 13.9 27.67 44.5 40.9 44.680.33 AD-11554 −12.6 −20.5 −32.1 −12.6 −17.7 −8.0 AD-11555 −20.8 −22.2−36.7 −9.8 −8.9 0.8 AD-11556 6.4 −11.6 −21.7 29.03 11.1 37.1 34.7 −17.37AD-11557 30.6 22.0 −8.7 −175.73 41.5 48.3 78.3 82.20 AD-11558 3.0 1.2−33.7 72.97 −23.5 −18.2 −30.8 82.49 AD-11559 −29.1 −38.0 −42.9 −2.4−37.3 −26.5 AD-11560 −42.0 −46.1 −54.6 19.8 0.7 19.1 AD-11561 −16.4−16.6 −15.9 37.38 50.5 61.2 57.4 −129.83 AD-11562 12.0 −13.9 −63.1 61.33−35.3 −46.2 −42.4 63.56 AD-11563 −45.3 −51.6 −61.7 −45.3 −54.7 −42.9AD-11564 −19.0 −31.3 −44.0 −38.5 −32.3 −0.1 AD-11565 −27.0 −4.4 −10.750.32 6.7 47.4 52.9 26.95 AD-11566 6.8 6.0 −0.5 32.0 18.2 0.7 AD-11567−1.1 −5.7 0.3 3.1 −4.6 14.4 AD-11568 10.3 7.9 11.4 −0.7 27.6 25.6AD-11569 17.6 22.0 15.4 33.01 47.1 51.3 63.7 −85.01 AD-11570 23.8 5.9−15.4 98.14 9.3 −5.5 −15.3 −217.80 AD-11571 −27.5 −23.9 −32.4 −8.5 −13.010.8 AD-11572 −35.2 −25.7 −26.1 4.3 8.8 18.6 AD-11573 −8.3 16.7 15.746.60 55.9 76.5 37.8 −22.05 AD-11574 1.1 −12.0 −27.6 12.5 −2.9 −11.3AD-11575 −29.9 −30.0 −35.9 1.6 27.3 −11.4 AD-11576 −9.4 −8.2 −20.8 0.134.6 3.1 AD-11577 −8.5 −0.1 13.7 45.2 31.5 23.2 AD-11578 9.8 17.0 4.1−40.1 −56.3 −53.3 AD-11579 15.1 12.2 −7.8 −47.6 −37.3 −18.5 AD-11580−4.4 2.3 4.1 76.38 14.2 4.9 33.6 12.97 AD-11581 10.6 2.5 3.4 −26.97 57.465.1 81.7 −109.94 AD-11582 10.6 4.3 −33.2 −1.17 −99.7 −93.3 −90.8−533.33 AD-11583 −16.6 −18.4 −28.3 −85.8 −72.4 −55.3 AD-11584 −24.1−12.0 −18.7 59.87 −1.0 54.0 16.5 1.61 AD-11585 −7.2 −9.6 9.1 51.46 37.519.1 45.7 82.71 AD-11586 −6.8 −0.7 −9.1 −81.1 −87.9 −86.9 AD-11587 6.1−0.4 0.9 −80.7 −6.2 −51.8 AD-11588 20.9 10.4 −9.5 99.40 −12.9 −0.1 20.5−117.23 AD-11589 28.7 24.3 16.1 35.75 18.2 38.7 54.2 88.14 AD-11590 20.728.8 3.0 98.81 −38.5 −55.2 20.9 44.92 AD-11591 −18.3 −29.6 −27.0 −3.1−40.1 −30.1 AD-11592 −11.3 −14.7 −12.3 −36.3 −38.7 −46.1 AD-11593 3.85.0 −12.3 1.8 −15.8 4.6 AD-11594 33.4 4.9 −3.2 99.07 −25.2 −31.3 −21.77.63 AD-11595 −30.9 −37.7 −54.2 −11.6 −2.7 −2.7 AD-11596 −8.2 −14.8 −3.3−7.8 26.8 3.5 AD-11597 13.0 13.0 11.7 21.7 28.2 25.2 83.00 AD-11598 40.0−1.6 −10.4 98.91 −30.2 −33.7 −17.7 0.00 AD-11599 13.0 24.6 −18.3 98.79−20.9 −22.8 −50.8 34.32 AD-11600 16.7 2.8 −8.1 98.52 −4.3 −28.5 6.8−120.34 AD-11601 10.4 25.3 2.0 71.20 23.0 6.3 7.9 −652.16 AD-11602 39.745.0 24.4 38.73 47.4 37.7 54.8 48.13 AD-11603 41.2 42.4 36.0 54.85 38.235.7 43.8 73.04 AD-11604 39.6 36.3 31.6 −1.51 37.0 56.6 52.8 84.65AD-11605 50.2 38.8 21.2 60.19 48.1 58.6 77.6 66.95 AD-11606 41.5 36.81.4 89.23 53.2 33.2 37.0 79.32 AD-11607 −12.8 2.7 −9.9 38.83 29.0 8.321.4 −13.98 AD-11608 10.9 −6.7 6.1 14.24 39.5 49.3 42.9 −75.07 AD-1160927.3 32.9 29.0 51.46 36.8 40.7 50.2 20.81 AD-11610 23.9 19.7 16.0 23.9537.7 70.3 49.6 97.93 AD-11611 5.6 11.9 16.1 30.7 8.5 15.6 AD-11612 −2.9−4.0 −14.4 9.9 21.9 10.0 AD-11613 13.4 32.7 23.7 −2.6 17.5 44.4 AD-116149.0 14.3 10.3 21.9 9.9 −29.6 AD-11615 −5.9 22.3 −0.9 −24.8 −16.9 −21.2AD-11616 2.3 5.5 11.0 −10.4 −7.6 26.4 AD-11617 14.3 5.3 −1.9 −617.1539.8 62.5 91.5 14.55 AD-11618 −2.5 −4.3 −1.2 −11.0 −26.8 −28.6 AD-11619−8.3 −37.8 −30.2 −33.9 −43.8 −23.9 AD-11620 11.0 −19.2 −37.5 −30.5 −12.00.7 AD-11621 −29.1 −4.5 4.7 −255.66 60.9 64.7 78.8 74.06 AD-11622 11.915.0 4.2 9.8 −29.1 −15.8 AD-11623 7.7 −17.8 −24.6 −27.3 −48.1 −19.4AD-11624 −16.3 −36.6 −45.8 −36.1 −17.2 −8.3 AD-11625 45.0 11.0 0.3 37.9819.5 48.2 43.1 −23.73 AD-11626 −3.6 13.2 11.9 −25.4 −81.4 −80.3 AD-1162734.1 24.7 21.1 48.69 −46.8 −73.0 −54.8 12.71 AD-11628 −8.9 −1.4 −13.89.60 −9.5 6.1 35.2 59.80 AD-11629 22.9 23.0 11.8 −73.11 65.3 71.2 75.13.39 AD-11630 12.2 19.0 −2.0 −99.9 −123.8 −85.0 AD-11631 −10.4 −15.2−18.3 −127.0 −87.2 −59.2 AD-11632 −50.7 −49.9 −49.4 −128.91 −32.4 1.329.8 51.76 AD-11633 −39.4 −10.2 1.8 −19.31 20.6 42.9 57.4 49.57 AD-116342.2 12.3 −14.1 −55.8 −129.3 −136.9 AD-11635 −44.3 −46.0 −29.5 −60.2−89.0 −66.2 AD-11636 −48.8 −44.7 −41.1 −35.4 −17.4 7.3 AD-11637 −36.8−27.2 −17.3 59.55 34.3 33.8 52.8 80.40 AD-11638 −8.4 8.5 −3.9 −9.4 12.428.9 AD-11639 −47.6 −1.7 36.0 23.31 2.9 20.2 12.6 −97.46 AD-11640 69.439.4 34.6 75.14 66.0 71.5 82.5 49.15 AD-11641 42.6 15.2 15.9 65.14 73.181.9 87.7 96.50 AD-11642 24.8 21.4 32.7 −42.93 89.8 87.4 87.4 4.03AD-11643 37.7 53.8 52.7 89.14 94.3 94.2 97.5 −22.03 AD-11644 54.1 17.75.5 94.29 26.7 38.5 51.2 31.78 AD-11645 5.9 2.5 −2.5 51.97 64.7 68.180.1 93.34 AD-11646 −1.7 2.1 7.9 59.55 72.6 76.8 79.0 92.72 AD-11647 8.48.8 37.5 57.28 75.6 81.7 93.7 74.71 AD-11648 56.9 19.2 8.1 66.10 −15.4−9.9 13.0 71.19 AD-11649 −3.3 −6.9 −3.5 65.47 36.0 54.2 64.9 82.84AD-11650 1.9 −2.5 −1.5 17.28 58.7 65.3 59.6 2.31 AD-11651 6.7 9.4 30.965.70 70.3 87.1 91.2 83.96 AD-11652 55.8 27.7 10.3 −56.8 59.6 68.4 33.90AD-11653 12.4 13.8 10.9 86.84 66.4 70.4 75.8 2.02 AD-11654 13.8 7.2 10.776.1 73.8 73.8 −123.92 AD-11655 9.1 14.6 40.5 −7.01 82.4 83.9 92.6−146.11 AD-11656 39.1 9.6 −1.0 81.71 −21.9 30.7 19.0 20.34 AD-11657 −1.11.1 0.2 −3.1 20.7 48.9 AD-11658 7.3 0.9 1.5 86.84 55.5 50.8 41.3 93.52AD-11659 2.9 4.1 28.0 78.79 46.4 51.1 66.6 79.47 AD-11660 23.8 13.0 0.980.57 11.0 20.8 −57.0 90.51 AD-11661 −6.4 −8.0 −6.1 −37.1 −16.9 19.1AD-11662 −1.6 −7.9 −7.9 10.9 10.7 27.7 AD-11663 1.6 5.6 28.1 25.4 20.430.8 AD-11664 11.9 7.6 3.9 50.16 54.4 63.1 66.8 77.95 AD-11665 5.1 1.20.2 15.10 52.7 65.2 62.3 −13.26 AD-11666 0.0 −3.5 5.3 5.72 67.9 80.380.3 79.54 AD-11667 3.7 2.9 8.0 47.73 83.6 80.4 90.6 84.70 AD-11668 16.3−0.3 −1.3 −91.6 −4.4 2.8 AD-11669 −1.0 3.8 0.3 3.4 33.5 43.3 AD-11670−0.1 −4.6 −2.6 35.28 42.7 43.6 62.6 −5.48 AD-11671 0.1 1.1 16.0 80.5861.0 65.1 72.7 13.54 AD-11672 8.2 −0.1 −2.2 −69.9 25.3 −57.8 AD-116731.7 0.9 3.2 −46.1 −22.7 1.5 AD-11674 −4.2 −3.0 −6.0 34.4 14.8 32.2AD-11675 −0.6 0.6 18.4 11.1 45.1 59.7 AD-11676 22.1 11.5 7.7 78.86 75.786.4 78.6 16.95 AD-11677 5.6 −3.1 3.2 86.1 78.0 87.1 1.27 AD-11678 −2.3−7.8 5.0 87.06 86.4 86.6 86.6 −141.35 AD-11679 5.6 7.3 34.4 28.26 88.388.6 90.9 −79.25 AD-11680 30.4 7.9 −0.5 75.62 −38.9 13.4 −88.3 72.03AD-11681 −3.7 −10.1 −0.2 14.4 25.0 24.2 AD-11682 −10.0 −11.0 −7.7 −39.953.8 57.2 AD-11683 −6.4 6.6 32.3 92.99 59.6 53.3 48.2 57.85 AD-1168431.5 14.4 2.9 33.26 −156.0 −119.6 −198.2 87.80 AD-11685 0.0 −6.5 −9.5−6.15 −43.0 −107.3 −93.7 −20.61 AD-11686 −12.0 −7.9 −4.7 88.12 −134.6−69.2 −151.8 43.11 AD-11687 −4.8 6.3 29.0 −77.4 −41.4 −79.0 AD-1168840.0 28.5 26.7 −15.66 73.4 79.8 88.0 66.95 AD-11689 34.4 27.1 32.4 57.3383.4 74.8 86.5 85.59 AD-11690 24.0 30.2 42.1 71.71 82.5 89.4 89.4 83.05AD-11691 47.9 44.1 55.3 87.43 92.0 93.9 97.0 70.96 AD-11694 44.3 8.5 5.72.1 4.8 13.6 AD-11695 1.5 0.5 −5.3 42.1 36.7 46.0 AD-11696 5.1 4.8 10.849.2 56.9 76.3 AD-11698 18.4 20.4 45.5 59.09 69.5 79.5 82.0 54.66AD-11700 30.4 8.5 −2.1 84.00 −151.2 −108.0 −67.5 4.52 AD-11704 −7.2 −6.1−9.9 −37.2 −56.5 −32.4 AD-11705 −2.9 −2.4 −4.7 −7.3 0.9 −14.8 AD-117068.5 21.1 99.9 21.5 −8.9 33.9 AD-11707 41.8 25.9 21.9 90.69 79.3 84.285.1 63.14 AD-11708 27.2 27.4 28.1 83.92 80.0 81.1 83.7 24.58 AD-1171031.0 37.7 43.0 77.54 86.9 89.6 89.6 99.70 AD-11711 33.8 36.5 48.2 73.4391.1 89.8 93.9 −34.75 AD-11712 81.1 20.2 8.0 79.71 22.6 −4.3 27.5 −13.56AD-11713 −8.6 −4.0 −2.1 39.1 39.1 51.8 AD-11714 −8.7 −8.3 −4.3 63.2745.0 66.1 74.5 39.48 AD-11715 0.8 14.9 40.7 −92.23 79.0 82.0 86.9 1.15AD-11716 32.4 8.3 1.8 83.05 −46.0 −51.4 −11.3 −577.97 AD-11717 2.2 −0.5−0.6 −1.5 −13.0 −14.3 AD-11718 −8.1 −11.9 −19.8 2.4 18.8 29.2 AD-117194.2 11.6 33.8 57.93 55.3 56.6 74.0 −2.02 AD-11720 29.8 17.3 10.4 77.0586.4 91.8 95.6 −373.73 AD-11721 14.6 13.7 12.4 82.56 90.8 91.1 95.1−31.70 AD-11722 17.0 15.1 23.7 82.96 94.1 91.7 91.7 −112.39 AD-1172316.0 13.5 30.5 90.99 94.1 95.5 95.7 −55.62 AD-11724 33.9 10.4 5.1 99.9863.2 73.2 73.7 −1.27 AD-11725 6.4 −2.0 0.0 93.62 85.5 92.4 89.5 −172.62AD-11726 2.4 0.2 0.2 58.58 90.3 91.9 94.4 −2.54 AD-11727 −1.0 2.8 20.24.72 93.9 94.5 98.3 49.86 AD-11728 30.7 12.9 4.0 73.43 26.9 27.5 41.030.51 AD-11729 5.1 1.5 −4.3 26.86 51.9 65.4 58.9 −20.46 AD-11730 2.6 1.30.8 24.70 60.2 67.7 71.6 52.02 AD-11731 4.9 3.3 24.1 79.94 66.8 67.389.0 96.16 AD-11732 33.6 9.3 6.0 63.43 63.0 79.9 87.5 70.34 AD-11733 2.22.8 −0.7 75.57 84.2 89.7 85.1 −9.51 AD-11734 8.6 6.7 10.8 90.52 90.890.6 90.6 −119.16 AD-11735 6.9 9.3 31.4 −22.87 90.8 88.5 94.5 −77.23AD-11736 31.5 9.9 3.0 21.14 29.9 39.5 45.3 −611.86 AD-11737 −8.0 −3.4−1.7 −183.17 78.3 88.4 82.5 14.41 AD-11738 −0.2 1.5 −4.7 −102.05 85.386.4 80.4 10.52 AD-11739 7.4 3.8 21.5 52.75 53.8 55.2 83.5 89.27AD-11740 30.0 11.7 −0.4 53.43 −87.3 −64.8 12.1 38.98 AD-11741 −3.5 −6.1−2.9 42.39 27.2 47.7 36.1 −32.56 AD-11742 −1.0 −1.9 −6.0 40.35 49.9 56.920.5 12.25 AD-11743 0.1 6.6 26.9 37.35 46.3 24.7 71.2 26.08 AD-1174440.5 24.2 21.0 75.24 42.9 52.0 90.3 32.20 AD-11745 26.3 23.1 23.3 55.4364.3 74.1 84.5 54.24 AD-11746 7.2 0.9 −4.8 89.82 42.3 74.8 8.6 33.49AD-11747 1.6 −3.5 −5.4 54.9 5.8 25.4 AD-11748 −4.3 0.5 0.9 30.3 10.910.9 AD-11749 8.4 3.9 1.0 41.4 −3.7 54.3 AD-11750 4.8 2.1 0.0 44.9 30.471.5 AD-11751 3.3 0.6 −2.2 34.3 −18.6 29.4 AD-11752 17.7 6.5 3.2 −27.6319.8 19.6 −0.1 61.08 AD-11753 1.7 1.2 0.9 −62.5 −40.7 28.2 AD-11754 3.71.8 0.7 69.9 91.1 44.9 AD-11755 37.7 26.6 10.8 −58.93 10.1 0.6 28.847.05 AD-11756 −1.0 −0.6 0.3 −39.8 −16.7 −5.7 AD-11757 0.2 0.0 1.8 −46.4−12.7 −2.7 AD-11758 18.9 11.7 7.5 21.83 27.3 12.4 −9.8 70.02 AD-1175926.1 15.1 18.8 −595.93 −6.9 −3.8 −22.6 55.98 AD-11760 21.9 19.5 6.011.60 −61.2 6.6 6.6 22.49 AD-11761 18.1 8.6 6.4 −67.30 39.7 43.4 58.071.77 AD-11762 15.4 9.7 4.2 −335.24 38.7 −86.0 12.4 73.68 AD-11763 6.87.3 3.7 −3.6 −0.7 8.7 AD-11764 5.1 5.7 0.9 −34.9 28.7 18.2 AD-11765 14.88.0 4.3 5.98 32.1 27.3 58.4 76.08 AD-11766 2.9 3.5 3.9 13.4 −6.1 −68.1AD-11767 24.1 15.1 5.5 7.18 32.9 −27.7 0.2 9.57 AD-11768 19.2 16.5 4.330.34 −92.5 −3.0 4.0 55.98 AD-11769 5.6 2.3 2.6 6.7 25.7 43.9 AD-1177014.0 7.2 2.2 −16.41 19.1 34.2 35.3 49.60 AD-11771 2.8 2.1 1.3 37.7 19.115.6 AD-11772 4.0 2.2 2.6 70.0 52.3 52.3 AD-11773 1.3 −0.6 −0.7 52.065.8 65.4 AD-11774 2.0 1.5 0.5 73.1 −18.3 6.5 AD-11775 2.3 0.3 −0.2 57.849.1 5.5 AD-11776 2.7 0.4 1.6 59.6 48.4 41.0 AD-11777 3.6 1.3 0.0 10.447.2 53.8 AD-11778 0.9 −0.9 −0.8 40.4 −13.5 −17.7 AD-11779 6.3 1.7 −0.542.1 2.9 −6.9 AD-11780 5.8 0.1 0.1 29.5 −17.6 20.3 AD-11781 2.3 1.0 −0.28.8 33.5 39.1 AD-11782 −1.7 −4.0 −4.1 48.1 44.5 56.1 AD-11783 −3.5 −2.1−1.5 34.8 16.7 44.2 AD-11784 5.7 3.7 −1.0 50.4 59.1 59.1 AD-11785 2.81.0 1.5 75.3 88.0 81.7 AD-11786 14.9 6.5 −2.9 97.46 −7.0 −34.8 −6.566.19 AD-11787 −0.3 0.0 −2.9 9.0 −8.6 18.2 AD-11788 −0.1 0.5 1.3 23.511.8 55.0 AD-11789 5.1 5.7 3.1 57.8 45.6 62.5 AD-11790 5.7 0.0 −4.0 18.8−12.3 −1.9 AD-11791 −1.5 −2.6 −4.6 42.1 −27.4 10.4 AD-11792 −0.9 1.8 1.527.1 30.8 24.6 AD-11793 2.2 3.7 4.4 6.7 34.8 44.3 AD-11794 0.4 −0.5 −1.659.5 81.9 69.3 AD-11795 −1.9 −0.8 0.1 78.7 94.4 87.2 AD-11796 5.0 1.80.8 89.6 71.8 71.8 AD-11797 0.6 0.8 1.3 92.1 87.0 83.7 AD-11798 −0.2−0.2 −1.8 67.4 69.7 45.4 AD-11799 −2.1 −1.9 −1.3 −49.1 −5.6 72.9AD-11800 1.3 −1.0 −0.4 64.1 58.5 60.6 AD-11801 0.1 0.1 0.8 23.7 35.354.5 AD-11802 4.8 0.8 −0.4 −115.9 −81.0 −113.4 AD-11803 3.7 1.3 −2.110.5 22.6 16.3 AD-11804 −2.0 −1.1 0.0 16.0 19.8 36.1 AD-11805 −0.5 0.2−0.4 −6.9 6.5 51.6 AD-11806 −11.1 −16.4 −13.0 79.5 70.3 29.3 AD-1180713.3 −13.2 −11.2 −78.63 65.7 20.0 50.5 −58.37 AD-11808 −6.3 −9.0 9.427.5 30.6 30.6 AD-11809 −0.2 −11.7 −11.7 44.7 61.3 73.0 AD-11810 −15.2−17.1 −17.8 80.1 81.6 81.7 AD-11811 −9.7 −12.1 −11.2 40.4 38.0 23.7AD-11812 −8.1 −12.5 −15.0 21.4 12.4 22.7 AD-11813 29.3 −14.5 −6.8−258.78 31.4 32.0 43.9 −423.13 AD-11814 −19.9 −18.8 −8.1 49.2 15.1 55.1AD-11815 −12.8 −13.4 −14.9 62.5 29.3 36.0 AD-11816 −14.5 −15.2 −14.613.0 −11.9 46.2 AD-11817 −15.4 −15.2 −11.8 43.3 38.2 42.0 AD-11818 9.5−1.9 −6.6 −176.59 69.0 43.6 35.2 39.23 AD-11819 −3.5 −1.4 −1.9 75.0 50.731.2 AD-11820 7.6 4.4 −2.0 −327.48 58.5 50.6 50.6 29.19 AD-11821 1.3−0.5 1.6 42.4 26.4 49.4 AD-11822 7.7 −4.1 −4.2 82.6 70.2 71.9 AD-11823−1.5 −2.3 −0.4 63.9 58.8 37.4 AD-11824 3.2 1.7 −2.7 33.5 13.3 37.3AD-11825 0.1 −0.9 0.7 53.6 42.2 44.8 AD-11826 3.4 −6.1 −3.8 52.8 35.739.9 AD-11827 −4.2 −2.6 −1.1 54.5 26.3 15.2 AD-11828 0.8 −1.3 −1.4 9.3−4.3 19.5 AD-11829 2.0 −1.6 1.1 32.2 16.0 32.0 AD-11830 −4.3 −7.5 −8.044.5 48.3 75.6 AD-11831 −6.4 −5.2 −4.1 39.3 56.5 67.3 AD-11832 −3.2 −2.5−3.0 61.0 76.2 76.2 AD-11833 −1.4 −2.5 −1.2 80.6 87.2 91.0 AD-11834 18.82.8 −7.7 −260.69 7.3 −11.5 24.5 25.84 AD-11835 −2.0 −0.2 −4.6 52.7 59.461.0 AD-11836 3.3 0.7 −1.9 70.7 76.0 80.0 AD-11837 0.3 −2.3 −0.7 84.670.1 80.8 AD-11838 −7.4 −7.1 −8.8 −13.5 9.5 8.9 AD-11839 −5.6 −2.9 −2.2−0.7 45.3 48.4 AD-11840 −1.7 1.3 −0.2 85.7 61.6 59.6 AD-11841 −3.8 −1.8−2.3 60.7 52.5 50.8 AD-11842 14.4 7.7 0.8 35.57 −145.5 −117.2 −46.442.11 AD-11843 ND ND ND −68.1 −87.6 −36.1 AD-11844 0.0 1.4 −0.5 −72.1−80.2 −80.2 AD-11845 3.9 2.0 1.1 −34.1 −60.1 −13.9 AD-11846 27.5 20.0−7.1 25.19 −69.7 −62.0 −63.7 13.40 AD-11847 −5.9 −4.8 −6.2 −93.2 −61.3−57.4 AD-11848 −1.3 0.6 −1.7 −89.3 −76.1 −18.3 AD-11849 −1.0 −1.4 50.6−0.9 12.6 18.7 AD-11850 −4.1 2.9 −6.3 −15.1 −3.7 −34.5 AD-11851 −5.0−4.6 −2.2 −13.3 −10.3 −15.2 AD-11852 −0.4 −0.6 −0.6 7.7 11.6 10.2AD-11853 1.6 −0.6 −0.8 24.9 28.1 36.7 AD-11854 −0.6 −1.3 −1.8 50.9 35.621.2 AD-11855 5.0 3.4 1.6 47.8 24.7 28.6 AD-11856 2.3 0.9 −0.6 60.4 36.736.7 AD-11857 3.4 0.7 0.1 23.1 37.0 57.6 AD-11858 −2.3 −2.5 −2.7 59.878.3 41.7 AD-11859 0.6 2.0 −1.0 64.0 33.3 40.7 AD-11860 11.4 6.4 0.132.44 40.2 −5.8 34.3 54.70 AD-11861 3.2 1.7 −0.4 −6.3 −30.9 26.8AD-11862 15.3 3.2 −3.1 −55.53 37.3 6.8 51.0 51.20 AD-11863 8.7 8.8 4.1−77.56 26.0 17.7 37.8 80.86 AD-11864 8.9 3.5 −1.4 −179.90 40.7 27.9 −8.969.38 AD-11865 4.3 0.2 0.1 0.3 8.6 14.9 AD-11870 4.5 5.7 1.9 60.1 43.268.8 AD-11871 13.9 4.6 −0.4 86.72 59.8 62.3 72.1 AD-11872 12.4 9.9 −0.281.93 75.1 77.9 77.9 79.43 AD-11873 16.2 8.7 2.9 51.91 81.9 90.3 92.0AD-11874 14.0 3.6 −0.6 63.74 36.6 41.1 21.2 AD-11875 37.1 22.5 9.9 92.1154.4 52.0 47.5 76.56 AD-11876 11.8 4.2 1.3 5.85 35.7 43.8 50.3 AD-118783.5 −0.5 −1.7 65.6 62.4 60.0 AD-11879 30.5 21.5 3.8 93.89 −4.3 −19.311.4 80.10 AD-11882 16.0 12.4 5.1 80.92 9.1 4.8 19.5 77.03 AD-11883 13.26.8 1.4 −37.40 −29.3 12.5 26.9 −1048.33 AD-11884 23.7 11.4 5.5 25.8341.4 25.2 40.9 −42.26 AD-11885 1.5 −0.8 −2.9 55.9 53.3 53.8 AD-11886−1.2 −3.4 −4.7 38.7 47.2 67.9 AD-11887 16.6 4.6 −2.2 −85.11 69.8 54.154.1 85.65 AD-11888 12.9 3.8 2.3 62.2 66.5 65.7 AD-11889 12.2 5.9 −1.64.6 10.5 40.1 AD-11890 8.5 6.2 5.0 −41.7 −12.9 20.9 AD-11891 20.7 9.81.0 −69.47 19.3 23.9 32.0 AD-11892 −0.1 −2.5 −2.0 38.4 32.1 38.0AD-11893 0.1 −2.1 −1.5 27.1 21.1 3.1 AD-11896 3.5 1.3 −1.6 23.3 −6.710.0 AD-11897 1.6 −1.3 −2.6 26.4 13.7 13.1 AD-11899 8.3 1.9 0.6 16.420.9 28.9 AD-11901 2.5 3.1 1.4 −89.3 71.9 78.6 AD-11902 5.9 0.6 −2.6−15.2 62.9 79.8 AD-11903 1.1 −1.5 −3.6 64.0 45.9 45.9 AD-11904 0.7 0.4−0.2 46.4 16.9 70.6 AD-11905 6.9 2.1 −0.1 −42.4 42.5 27.9 AD-11906 −0.40.3 −2.5 98.5 93.5 54.3 AD-11907 4.5 0.5 −1.1 95.8 45.3 95.1 AD-11908−0.9 −1.7 0.2 39.0 86.6 84.5 AD-11909 7.0 2.9 1.1 79.1 −27.1 83.9AD-11911 5.4 2.6 −1.9 97.4 98.6 94.0 AD-11912 −3.4 −4.5 −3.7 99.4 96.265.9 AD-11914 −0.6 −0.2 1.0 91.7 69.5 91.5 AD-11918 −6.3 −8.2 −6.7 −12.49.7 8.0 AD-11919 −1.0 −0.1 3.2 18.2 −9.2 32.5 AD-11925 3.5 1.9 −2.7 11.913.7 13.7 AD-11926 11.3 4.9 −3.2 46.95 16.9 37.0 66.5 93.16 AD-11927−6.8 −8.4 −5.7 45.5 −12.2 −17.2 AD-11933 −0.2 1.4 3.3 −6.7 32.7 3.5AD-11938 0.0 −2.4 −4.1 19.2 −45.8 1.4 AD-11939 −2.3 −2.0 −3.1 3.0 23.124.9 AD-11941 −8.3 −9.6 −7.5 42.1 4.0 −25.0 AD-11942 −4.4 −3.5 −2.5 31.62.5 1.4 AD-11943 1.9 0.7 −2.7 −5.6 20.5 15.4 AD-11944 −1.1 −2.1 −2.051.0 46.7 50.6 AD-11945 −7.7 −8.6 −5.9 77.2 30.5 55.8 AD-11946 0.6 0.8−2.4 66.9 65.2 65.4 AD-11947 −0.6 −2.2 −4.1 66.6 68.5 68.5 AD-11948 −2.3−4.0 −4.5 77.1 77.6 88.6 AD-11949 −6.6 −7.9 −6.4 67.5 33.7 2.9 AD-11950−3.9 0.8 7.9 56.0 51.7 44.8 AD-11951 7.7 0.4 6.0 35.6 45.6 46.1 AD-119521.7 −4.5 −4.7 54.8 65.7 66.0 AD-11953 −9.1 −7.7 −6.8 51.4 22.8 30.1AD-11954 −2.7 8.2 8.2 42.1 26.4 32.1 AD-11955 9.0 7.5 5.9 54.6 43.4 44.2AD-11956 6.3 −6.5 −4.0 28.1 40.0 55.3 AD-11957 −1.9 −6.4 −2.8 78.2 95.969.2 AD-11958 2.0 1.2 −1.0 99.7 89.5 86.2 AD-11959 8.7 3.4 −1.2 81.274.5 74.5 AD-11960 2.8 1.7 0.2 71.9 61.4 82.7 AD-11961 −3.4 −5.5 −4.584.3 50.6 68.7 AD-11962 −1.4 0.0 −0.2 74.1 89.2 63.9 AD-11963 2.1 −0.3−0.2 78.8 20.1 50.9 AD-11964 4.5 3.9 −0.1 61.5 51.0 58.4 AD-11965 −3.8−6.3 −4.4 64.6 14.2 89.4 AD-11966 −0.5 −1.0 −0.5 49.5 −30.0 21.4AD-11967 5.0 3.8 1.2 24.7 28.2 24.2 AD-11968 −0.8 −0.2 −0.3 9.5 6.5 26.5AD-11969 3.9 0.6 −2.2 63.5 31.6 56.5 AD-11970 9.2 5.5 −1.6 47.6 36.957.1 AD-11971 −1.2 0.1 −1.7 63.8 55.3 55.3 AD-11972 2.7 −0.9 −0.8 66.782.1 78.1 AD-11973 3.2 5.3 −0.3 −137.3 −57.5 −69.7 AD-11974 1.7 2.3 2.6−15.1 −41.3 −55.7 AD-11975 1.8 1.4 1.9 −63.6 −63.1 −54.6 AD-11976 2.32.7 2.7 −8.3 10.1 29.0 AD-11977 4.5 3.7 2.7 −168.8 −142.8 −116.4AD-11978 2.1 1.2 0.8 −66.5 −91.0 −120.2 AD-11979 −1.7 −2.0 −2.2 −140.3−128.4 −107.4 AD-11980 −1.3 −0.8 −0.6 −21.5 −10.2 −14.2 AD-11981 9.1 6.74.6 −181.1 −162.1 −164.3 AD-11982 −0.2 −0.4 −1.4 −189.5 −148.6 −159.6AD-11983 −2.7 −3.4 −3.7 −131.3 −111.7 −115.3 AD-11984 −1.9 −0.5 0.1−36.1 −12.9 −10.8 AD-11985 5.5 6.4 2.8 −139.4 −160.3 −216.6 AD-11986 6.16.0 5.8 −111.7 −143.8 −134.7 AD-11987 0.9 1.5 2.0 −162.1 −148.5 −132.2AD-11988 7.4 8.5 8.9 −196.2 −201.7 −178.2 AD-11989 1.9 2.0 1.6 −62.4−57.6 −61.2 AD-11990 0.8 −0.2 −0.8 −51.3 −69.3 −76.8 AD-11991 −1.9 −2.4−2.3 −84.5 −65.2 −51.0 AD-11992 0.6 1.3 1.6 −83.1 −60.4 −57.1 AD-119936.6 5.6 3.7 −29.9 −33.8 −35.4 AD-11994 0.3 0.1 −0.6 −60.1 −59.5 −59.4AD-11995 −2.3 −2.1 −1.9 −73.2 −61.7 −58.5 AD-11996 −0.7 0.3 0.4 −4.6−14.0 −21.4 AD-11997 4.1 4.3 −0.1 64.7 26.4 52.9 AD-11998 0.3 0.0 −0.769.1 61.0 52.1 AD-11999 −0.5 −0.1 0.1 31.9 37.3 39.5 AD-12000 −0.8 0.20.8 51.7 54.7 60.6 AD-12001 3.7 3.3 2.9 54.7 49.4 47.3 AD-12002 3.8 2.92.2 34.0 49.3 62.0 AD-12003 −0.3 −0.4 −0.2 26.3 29.4 32.9 AD-12004 0.60.8 1.4 37.7 36.6 36.9 AD-12005 4.9 4.7 3.9 19.6 9.3 7.5 AD-12006 0.80.5 0.5 −22.4 −23.6 −19.4 AD-12007 −1.3 −0.7 −0.3 −10.4 −13.6 −21.7AD-12008 0.7 1.4 1.4 31.6 31.4 33.4 AD-12009 0.8 2.5 −2.2 36.9 25.4 31.0AD-12010 −1.2 −1.5 −1.6 35.6 23.9 15.8 AD-12011 −1.2 −0.3 0.2 50.4 46.041.7 AD-12012 0.7 0.7 0.5 47.1 49.2 51.5 AD-12013 1.3 1.1 −0.5 31.5 25.221.8 AD-12014 −1.8 −3.0 −2.7 21.5 26.1 26.3 AD-12015 −4.2 −4.1 −3.8 36.535.7 34.2 AD-12016 −1.9 −1.5 −1.2 38.7 41.0 41.9 AD-12017 −0.4 −1.0 −2.26.4 12.0 17.5 AD-12018 −1.0 −2.6 −2.9 −19.0 −8.5 −7.1 AD-12019 −3.1 −2.7−2.3 −3.4 −3.2 0.0 AD-12020 −2.1 −1.4 −1.0 2.2 10.2 12.7 AD-12021 4.93.3 2.1 −34.2 −9.2 −30.6 AD-12022 3.9 3.1 2.1 −139.2 −184.9 −191.4AD-12023 0.4 0.5 0.8 10.7 11.7 −2.8 AD-12024 0.2 1.4 2.3 −8.4 6.0 18.1AD-12025 6.9 4.8 3.2 18.0 −1.4 −25.4 AD-12026 −0.3 −0.1 0.2 −79.8 −64.2−44.8 AD-12027 −0.8 −1.0 −0.5 −28.2 −43.7 −57.1 AD-12028 −1.2 −0.2 0.8−8.1 −0.6 6.2 AD-12029 6.9 5.6 3.8 −65.2 −79.3 −106.3 AD-12030 −0.1 −0.8−2.0 −70.4 −93.2 −119.7 AD-12031 0.2 0.1 −0.1 −32.4 −34.4 −34.9 AD-120320.0 0.1 0.6 −86.6 −68.5 −43.4 AD-12033 −4.7 −8.2 −7.6 −115.7 −149.7−58.5 AD-12034 −6.0 −5.7 −5.7 −217.6 −210.8 −215.6 AD-12035 −7.7 −7.4−7.6 −99.6 −102.4 −107.1 AD-12036 −6.6 −6.1 −5.1 −111.1 −91.0 −89.8AD-12037 −7.8 −7.8 −7.7 45.1 −5.3 −44.2 AD-12462 −8.4 −8.6 −9.4 −150.2−135.8 −130.8 AD-12463 −8.3 −8.4 −8.6 −70.1 −70.9 −75.8 AD-12464 −6.3−6.6 −6.5 −38.9 −19.7 −2.6 AD-12465 −2.8 −3.8 −5.3 −52.8 −75.2 −92.9AD-12466 −1.9 −3.1 −3.5 −68.5 −103.9 −113.9 AD-12467 −5.9 −5.8 −5.6−118.6 −121.8 −117.7 AD-12468 −4.9 −4.2 −3.3 −14.2 −8.4 −6.8 AD-12469−4.8 −6.0 −6.6 −29.1 −6.1 −10.2 AD-12470 −0.9 −1.4 −1.9 −39.5 −19.2 −1.8AD-12471 −4.0 −4.0 −4.0 −15.4 −16.3 −17.3 AD-12472 −3.2 −3.4 −2.6 25.924.0 27.4 AD-12473 −3.5 −3.8 −4.1 −42.5 −31.5 −26.4 AD-12474 −5.0 −5.3−5.8 −7.1 −3.0 −1.8 AD-12475 −7.1 −7.6 −7.4 −10.2 −4.3 −0.7 AD-12476−5.2 −4.7 −3.5 24.3 26.9 32.2 AD-12477 −2.8 −3.8 −4.4 −41.4 −45.4 −37.6AD-12478 −7.3 −7.6 −7.8 −59.7 −63.8 −67.5 AD-12479 −7.9 −8.3 −8.1 −54.9−39.3 −8.6 AD-12480 −2.7 −2.4 −1.4 9.3 18.0 12.4 AD-12481 −5.7 −5.5 −7.5−131.4 −152.8 −48.2 AD-12482 −4.4 −4.5 −4.1 −159.1 −127.8 −140.7AD-12483 −4.8 −4.8 −4.5 −66.0 −64.8 −60.3 AD-12484 −4.2 −3.9 −3.8 17.927.6 29.1 AD-12485 −2.7 −4.0 −5.4 −170.2 −176.6 −186.0 AD-12486 −8.5−8.6 −8.6 −200.1 −194.6 −169.0 AD-12487 −8.1 −8.0 −7.8 −117.5 −106.8−86.5 AD-12488 −8.2 −7.9 −7.8 −17.3 −13.1 0.8 AD-12489 −4.2 −5.6 −7.1−128.0 −152.3 −185.8 AD-12490 −8.6 −8.9 −8.8 −241.7 −258.1 −251.4AD-12491 −8.9 −9.0 −8.4 −181.6 −184.9 −188.0 AD-12492 −8.6 −7.7 −6.7−23.3 −22.4 −27.3 AD-12493 −8.2 −6.7 −4.5 −93.8 −54.8 6.7 AD-12494 −7.6−8.0 −8.2 −62.2 −63.5 −54.7 AD-12495 −6.6 −5.5 −4.9 16.6 0.4 −10.0AD-12496 −4.1 −4.2 −3.8 40.2 41.7 39.5 AD-12497 −5.1 −5.2 −6.3 −67.9−65.2 −67.3 AD-12498 −5.8 −6.0 −6.4 −68.6 −84.1 −94.1 AD-12499 −6.6 −6.3−6.3 −93.4 −80.2 −61.0 AD-12500 −6.0 −5.4 −4.7 9.7 10.2 11.3 AD-12501−4.8 −5.0 −5.8 −87.1 −108.5 −110.6 AD-12502 −6.8 −7.2 −6.7 −126.5 −125.5−119.1 AD-12503 −10.4 −9.6 −9.9 −85.2 −76.4 −86.7 AD-12504 −4.0 −3.6−2.5 −59.7 −52.7 −34.7 AD-12505 18.1 −0.9 −5.5 42.18 3.5 −116.5 −34.2−282.78 AD-12506 −2.1 −2.7 −4.2 −29.8 −22.3 −11.9 AD-12507 −1.7 7.2 15.044.47 29.1 33.0 32.3 AD-12508 −4.2 −3.2 −1.9 40.7 33.9 30.5 AD-12509−9.2 −8.9 −8.2 −54.3 −67.7 −80.3 AD-12510 −7.9 −7.6 −7.6 −127.6 −117.5−124.2 AD-12511 −7.1 −6.9 −6.8 −28.4 −18.6 −6.7 AD-12512 −5.2 −4.1 −3.3−0.4 −1.7 −1.5 AD-12513 −6.9 −7.4 −8.8 −65.5 −80.0 −100.6 AD-12514 −10.8−11.0 −10.6 −83.8 −85.0 −92.0 AD-12515 −10.7 −9.6 −9.3 −19.3 −41.2 −32.7AD-12516 −5.8 −5.3 −4.2 −5.9 0.9 15.9 AD-12517 −8.6 −8.7 −7.1 29.2 53.049.4 AD-12518 −8.3 −8.0 −8.0 65.7 57.8 53.6 AD-12519 −7.2 −7.2 −7.6 51.251.3 53.9 AD-12520 −7.0 −6.3 −5.4 67.7 73.2 78.8 AD-12521 −6.4 −8.0 −9.119.8 27.6 35.5 AD-12522 −9.9 −9.0 −9.1 50.3 43.0 34.3 AD-12523 −9.0 −8.7−8.9 28.3 31.9 38.1 AD-12524 −7.0 −7.4 −7.0 63.7 67.1 65.9 AD-12525 −1.6−3.8 −6.0 16.1 21.8 21.2

TABLE 4 In vitro Plaque assay controls (all values average of 3-4experiments and expressed as % inhibition relative to no siRNAtreatment) AD-1955 (luc) 31.37 10.97 AD-5179 (GFP) 15.15 18.65 LS L#177.23 N/A LS NP#1 73.94 N/A LS VP35#1 67.27 N/A

TABLE 5 Exo + endo seq parent light id seq duplex duplex Target senseantisense no sense 5′-3′ id no antisense 5′-3′ AD-11594 AD-3542 NP A-A-30769 995 cAGuAGGAcAcAuGAuGGudTsdT 996 ACcAUcAUGUGUCCuACUGdTsdT 30768AD-11596 AD-3543 NP A- A-30771 997 GuAGGAcAcAuGAuGGuGAdTsdT 998UcACcAUcAUGUGUCCuACdTsdT 30770 AD-11597 AD-3544 NP A- A-30773 999uAGGAcAcAuGAuGGuGAudTsdT 1000 AUcACcAUcAUGUGUCCuAdTsdT 30772 AD-11598AD-3545 NP A- A-30775 1001 GAuGGuGAuuuuccGuuuGdTsdT 1002cAAACGGAAAAUcACcAUCdTsdT 30774 AD-11600 AD-3546 NP A- A-30777 1003uGGuGAuuuuccGuuuGAudTsdT 1004 AUcAAACGGAAAAUcACcAdTsdT 30776 AD-11603AD-3547 NP A- A-30779 1005 cuGAGAAGcAAcuccAAcAdTsdT 1006UGUUGGAGUUGCUUCUcAGdTsdT 30778 AD-11604 AD-3548 NP A- A-30781 1007uGAGAAGcAAcuccAAcAAdTsdT 1008 UUGUUGGAGUUGCUUCUcAdTsdT 30780 AD-11606AD-3549 NP A- A-30783 1009 AGAAGcAAcuccAAcAAuAdTsdT 1010uAUUGUUGGAGUUGCUUCUdTsdT 30782 AD-11609 AD-3550 VP35 A- A-30785 1011AAGuGAuGAAGAuuAAGAAdTsdT 1012 UUCUuAAUCUUcAUcACUUdTsdT 30784 AD-11610AD-3551 VP35 A- A-30787 1013 AGuGAuGAAGAuuAAGAAAdTsdT 1014UUUCUuAAUCUUcAUcACUdTsdT 30786 AD-11611 AD-3552 VP40 A- A-30789 1015cuGccuGcuGcAAcAuGGAdTsdT 1016 UCcAUGUUGcAGcAGGcAGdTsdT 30788 AD-11613AD-3553 GP A- A-30791 1017 GGcuGAAAAcuGcuAcAAudTsdT 1018AUUGuAGcAGUUUUcAGCCdTsdT 30790 AD-11614 AD-3554 GP A- A-30793 1051GcuGAAAAcuGcuAcAAucdTsdT 1052 GAUUGuAGcAGUUUUcAGCdTsdT 30792 AD-11615AD-3555 GP A- A-30795 1053 cuGAAAAcuGcuAcAAucudTsdT 1054AGAUUGuAGcAGUUUUcAGdTsdT 30794 AD-11618 AD-3556 GP A- A-30797 1055AAAAcuGcuAcAAucuuGAdTsdT 1056 UcAAGAUUGuAGcAGUUUUdTsdT 30796 AD-11621AD-3557 GP A- A-30799 1057 AcuGcuAcAAucuuGAAAudTsdT 1058AUUUcAAGAUUGuAGcAGUdTsdT 30798 AD-11622 AD-3558 VP30 A- A-30801 1059AGcAAAuccAAcGGcuGAudTsdT 1060 AUcAGCCGUUGGAUUUGCUdTsdT 30800 AD-11624AD-3559 VP30 A- A-30803 1061 cAAAuccAAcGGcuGAuGAdTsdT 1062UcAUcAGCCGUUGGAUUUGdTsdT 30802 AD-11627 AD-3560 L A- A-30805 1063AuGcAuGucAGuGAuuAuudTsdT 1064 AAuAAUcACUGAcAUGcAUdTsdT 30804 AD-11628AD-3561 L A- A-30807 1065 uGcAuGucAGuGAuuAuuAdTsdT 1066uAAuAAUcACUGAcAUGcAdTsdT 30806 AD-11629 AD-3562 L A- A-30809 1067GcAuGucAGuGAuuAuuAudTsdT 1068 AuAAuAAUcACUGAcAUGCdTsdT 30808 AD-11630AD-3563 L A- A-30811 1069 cAuGucAGuGAuuAuuAuAdTsdT 1070uAuAAuAAUcACUGAcAUGdTsdT 30810 AD-11631 AD-3564 L A- A-30813 1071AuGucAGuGAuuAuuAuAAdTsdT 1072 UuAuAAuAAUcACUGAcAUdTsdT 30812

TABLE 6 In vitro plaque In vitro plaque In vitro assay against assayagainst plasmid Ebola-Zaire Ebola-Sudan modified screen IC50 (%inhibition (% inhibition duplex (nM) relative to no siRNA) relative tono siRNA) AD-3542 1.60 84% 74% AD-3543 0.00 74% 77% AD-3544 0.61 65% 13%AD-3545 0.00 82% 61% AD-3546 1.35 88% 71% AD-3547 0.00 81% 26% AD-35487.82 68% 73% AD-3549 0.00 68% 74% AD-3550 0.17 77% 22% AD-3551 0.08 2%75% AD-3552 0.00 78% 70% AD-3553 0.19 85% 26% AD-3554 0.77 84% 15%AD-3555 0.88 −5% 19% AD-3556 0.00 73% 74% AD-3557 0.21 100% 28% AD-35580.00 −7% 68% AD-3559 0.00 −252% −7% AD-3560 0.50 16% −35% AD-3561 0.00−105% 19% AD-3562 0.00 −75% 63% AD-3563 5.42 73% −17% AD-3564 0.97 −86%19% AD-3621 0.00 80% 48% AD-3622 16.00 61% 47% AD-3623 0.00 98% 37%AD-3624 0.00 84% 24% AD-3625 0.00 −7% −5% AD-3626 0.00 53% 31%

TABLE 7 IFN induction normalized TNF induction normalized to positivecontrol siRNA to positive control siRNA duplex % of control duplex % ofcontrol AD-11546 131.6 AD-11546 30.1 AD-11558 98.8 AD-11558 57.2AD-11570 0 AD-11570 0 AD-11588 147 AD-11588 0.0 AD-11590 0.0 AD-11590130.5 AD-11594 17.8 AD-11594 124.8 AD-11597 22.0 AD-11597 242.9 AD-1159825.7 AD-11598 180.3 AD-11599 136 AD-11599 183.9 AD-11600 13.5 AD-11600141.0 AD-11603 69.0 AD-11603 81.4 AD-11606 33.2 AD-11606 79.8 AD-11609126.9 AD-11609 46.3 AD-11610 138.2 AD-11610 48.5 AD-11611 43.2 AD-1161141.0 AD-11613 167.0 AD-11613 52.6 AD-11614 162.1 AD-11614 48.1 AD-11615171.2 AD-11615 60.0 AD-11618 137.3 AD-11618 54.6 AD-11621 0.0 AD-11621325.8 AD-11622 37.2 AD-11622 28.5 AD-11623 58.0 AD-11623 26.0 AD-1162463.2 AD-11624 29.2 AD-11627 6.4 AD-11627 125.0 AD-11628 0.0 AD-11628101.1 AD-11630 0.0 AD-11630 170.3 AD-11631 0.0 AD-11631 156.0 AD-11644 0AD-11644 0.0 AD-11650 0.0 AD-11650 0.0 AD-11659 0.0 AD-11659 69.0AD-11673 9.2 AD-11673 0.0 AD-11678 0.0 AD-11678 0.0 AD-11683 0.0AD-11683 19.6 AD-11684 0.0 AD-11684 24.3 AD-11691 0 AD-11691 0.0AD-11695 0.0 AD-11695 5.2 AD-11698 5.3 AD-11698 14.0 AD-11706 0.0AD-11706 0.0 AD-11707 0 AD-11707 0.0 AD-11710 0 AD-11710 0.0 AD-117210.0 AD-11721 0.0 AD-11725 0.0 AD-11725 0.0 AD-11732 0.0 AD-11732 0.0AD-11743 16.1 AD-11743 0.0 AD-11756 0.0 AD-11756 0.0 AD-11757 0.0AD-11757 0.0 AD-11758 0.0 AD-11758 0.0 AD-11759 0.0 AD-11759 0.0AD-11773 0.0 AD-11773 0.0 AD-11780 0.0 AD-11780 7.9 AD-11789 0.0AD-11789 5.7 AD-11804 0.0 AD-11804 0.0 AD-11811 0.0 AD-11811 0.0AD-11814 0.0 AD-11814 0.0 AD-11816 0.0 AD-11816 0.0 AD-11822 9.8AD-11822 0.0 AD-11823 0.0 AD-11823 0.0 AD-11832 0.0 AD-11832 0.0AD-11836 0.0 AD-11836 0.0 AD-11939 0.0 AD-11939 0.0 AD-11976 0.0AD-11976 0.0 AD-11982 6.9 AD-11982 15.1 AD-11990 12.2 AD-11990 0.0AD-11992 7.3 AD-11992 0.0 AD-12007 0.0 AD-12007 0.0 AD-12013 0.0AD-12013 0.0 AD-12019 24.6 AD-12019 5.0 AD-12024 0.0 AD-12024 19.8AD-12035 0.0 AD-12035 8.5 AD-12475 0.0 AD-12475 0.0 AD-12484 21.0AD-12484 9.5 AD-12491 0.0 AD-12491 13.1 AD-12500 12.6 AD-12500 53.3AD-12502 101.6 AD-12502 55.5 AD-3542 0.0 AD-3542 0.0 AD-3543 0.0 AD-35430.0 AD-3544 10.1 AD-3544 0.0 AD-3545 11.8 AD-3545 0.0 AD-3546 0.0AD-3546 0.0 AD-3547 0.0 AD-3547 31.0 AD-3548 0.0 AD-3548 5.9 AD-3549 0.0AD-3549 10.8 AD-3550 7.4 AD-3550 0.0 AD-3551 0.0 AD-3551 0.0 AD-3552 0.0AD-3552 0.0 AD-3553 0.0 AD-3553 0.0 AD-3554 0.0 AD-3554 11.5 AD-3555 0.0AD-3555 6.2 AD-3556 0.0 AD-3556 9.1 AD-3557 0.0 AD-3557 0.0 AD-3558 0.0AD-3558 5.8 AD-3559 0.0 AD-3559 5.3 AD-3560 0.0 AD-3560 0.0 AD-3561 0.0AD-3561 0.0 AD-3562 0.0 AD-3562 0.0 AD-3563 0.0 AD-3563 0.0 AD-3564 0.0AD-3564 0.0 AD-3621 0.0 AD-3621 0.0 AD-3622 0.0 AD-3622 0.0 AD-3623 5.7AD-3623 0.0 AD-3624 10.0 AD-3624 0.0 AD-3625 0.0 AD-3625 0.0 AD-3626 0.0AD-3626 0.0

TABLE 8 In vitro plasmid In vitro screen plasmid single dose screenduplex (% IC50 name silencing) (nM) AD-11542 9% AD-11543 −3% AD-11544−1% AD-11545 13% AD-11546 68% 0.72 AD-11547 28% AD-11548 48% AD-1154943% AD-11550 −5% AD-11551 8% AD-11552 −17% AD-11553 −6% AD-11554 15%AD-11555 2% AD-11556 6% AD-11557 −6% AD-11558 70% 0.46 AD-11559 28%AD-11560 5% AD-11561 30% AD-11562 24% AD-11563 6% AD-11564 0% AD-11565−4% AD-11566 4% AD-11567 −2% AD-11568 0% AD-11569 12% AD-11570 73% 0.95AD-11571 −2% AD-11572 −3% AD-11573 2% AD-11574 15% AD-11575 −2% AD-11576−9% AD-11577 −9% AD-11578 77% AD-11579 37% AD-11580 35% AD-11581 60%AD-11582 21% AD-11583 47% AD-11584 0% AD-11585 −1% AD-11586 36% AD-1158766% AD-11588 47% AD-11589 77% AD-11590 83% 0.57 AD-11591 65% AD-1159262% AD-11593 55% AD-11594 85% 0.35 AD-11595 72% AD-11596 84% 0.24AD-11597 85% 0.33 AD-11598 87% 0.21 AD-11599 91% 0.81 AD-11600 89% 0.29AD-11601 84% 1.07 AD-11602 71% AD-11603 80% 1.3 AD-11604 81% 1.44AD-11605 75% AD-11606 78% 6.38 AD-11607 53% AD-11608 60% AD-11609 75%0.3 AD-11610 74% 0.15 AD-11611 61% 0.28 AD-11612 −5% AD-11613 84% 0.077AD-11614 85% 0.102 AD-11615 79% 0.211 AD-11616 66% AD-11617 59% AD-1161878% 0.24 AD-11619 57% AD-11620 64% AD-11621 74% 0.15 AD-11622 70% 0.41AD-11623 67% 0.54 AD-11624 75% 0.15 AD-11625 11% AD-11626 51% AD-1162771% 0.28 AD-11628 68% 0.33 AD-11629 75% 0.18 AD-11630 73% 0.24 AD-1163169% 0.31 AD-11632 53% AD-11633 63% 1.78 AD-11634 65% 0.76 AD-11635 29%AD-11636 43% AD-11637 −5% AD-11638 6% AD-11639 2% AD-11640 38% AD-1164135% AD-11642 55% AD-11643 33% AD-11644 36% AD-11645 45% AD-11646 37%AD-11647 41% AD-11648 61% AD-11649 35% AD-11650 84% 0.7 AD-11651 13%AD-11652 64% AD-11653 61% AD-11654 6% AD-11655 59% AD-11656 38% AD-1165739% AD-11658 59% AD-11659 82% 0.038 AD-11660 39% AD-11661 −5% AD-11662−1% AD-11663 14% AD-11664 19% AD-11665 7% AD-11666 −4% AD-11667 −14%AD-11668 63% AD-11669 28% AD-11670 23% AD-11671 23% AD-11672 15%AD-11673 79% 0.117 AD-11674 67% AD-11675 46% AD-11676 20% AD-11677 34%AD-11678 79% 0.149 AD-11679 51% AD-11680 24% AD-11681 72% AD-11682 73%AD-11683 88% 0.056 AD-11684 80% 0.184 AD-11685 33% AD-11686 72% AD-1168732% AD-11688 15% AD-11689 58% AD-11690 26% AD-11691 60% AD-11694 54%AD-11695 81% 0.46 AD-11696 32% AD-11698 73% 0.2 AD-11700 −9% AD-1170435% AD-11705 39% AD-11706 67% 0.56 AD-11707 2% AD-11708 −10% AD-1171065% 4.57 AD-11711 −3% AD-11712 17% AD-11713 0% AD-11714 42% AD-11715 41%AD-11716 18% AD-11717 32% AD-11718 −3% AD-11719 36% AD-11720 41%AD-11721 68% 1.35 AD-11722 31% AD-11723 49% AD-11724 27% AD-11725 67%0.34 AD-11726 12% AD-11727 3% AD-11728 5% AD-11729 12% AD-11730 6%AD-11731 63% 49.4 AD-11732 76% 2.88 AD-11733 60% 8.76 AD-11734 44%AD-11735 17% AD-11736 44% AD-11737 14% AD-11738 −9% AD-11739 23%AD-11740 1% AD-11741 9% AD-11742 40% AD-11743 77% 0.11 AD-11744 24%AD-11745 27% AD-11746 −9% AD-11747 16% AD-11748 8% AD-11749 AD-11750 33%AD-11751 19% AD-11752 61% 6.92 AD-11753 13% AD-11754 53% AD-11755AD-11756 61% 1.21 AD-11757 63% 1.57 AD-11758 28% AD-11759 66% 1.61AD-11760 64% 3.4 AD-11761 59% AD-11762 54% AD-11763 47% AD-11764 21%AD-11765 −1% AD-11766 −1% AD-11767 67% 4.4 AD-11768 52% AD-11769 21%AD-11770 55% AD-11771 36% AD-11772 41% AD-11773 76% 0.37 AD-11774 35%AD-11775 49% AD-11776 50% AD-11777 −5% AD-11778 18% AD-11779 15%AD-11780 62% 0.76 AD-11781 14% AD-11782 38% AD-11783 46% AD-11784 23%AD-11785 −11% AD-11786 42% AD-11787 48% AD-11788 19% AD-11789 64% 0.38AD-11790 26% AD-11791 22% AD-11792 −8% AD-11793 26% AD-11794 57%AD-11795 AD-11796 59% AD-11797 11% AD-11798 11% AD-11799 35% AD-11800 2%AD-11801 −6% AD-11802 0% AD-11803 5% AD-11804 88% 0.281 AD-11805 5%AD-11806 9% AD-11807 6% AD-11808 50% AD-11809 24% AD-11810 −1% AD-1181166% 1.56 AD-11812 1% AD-11813 17% AD-11814 65% 0.43 AD-11815 −1%AD-11816 65% 0.99 AD-11817 67% 2.98 AD-11818 44% AD-11819 64% AD-1182036% AD-11821 64% AD-11822 62% 1.44 AD-11823 69% 0.32 AD-11824 38%AD-11825 18% AD-11826 23% AD-11827 2% AD-11828 51% AD-11829 AD-11830 46%AD-11831 20% AD-11832 71% 0.94 AD-11833 3% AD-11834 23% AD-11835 −8%AD-11836 65% 1.46 AD-11837 26% AD-11838 −16% AD-11839 22% AD-11840 −5%AD-11841 60% AD-11842 19% AD-11843 57% AD-11844 18% AD-11845 8% AD-1184648% AD-11847 57% AD-11848 2% AD-11849 66% 0.32 AD-11850 25% AD-11851−12% AD-11852 27% AD-11853 28% AD-11854 55% AD-11855 43% AD-11856 −20%AD-11857 61% 3.28 AD-11858 6% AD-11859 4% AD-11860 79% AD-11861 31%AD-11862 48% AD-11863 85% 0.39 AD-11864 42% AD-11865 37% AD-11870 34%AD-11871 79% 0.63 AD-11872 70% AD-11873 70% AD-11874 39% AD-11875 39%AD-11876 34% AD-11878 −1% AD-11879 50% AD-11882 −6% AD-11883 11%AD-11884 7% AD-11885 −3% AD-11886 −5% AD-11887 18% AD-11888 41% AD-118891% AD-11890 44% AD-11891 20% AD-11892 37% AD-11893 29% AD-11896 1%AD-11897 41% AD-11899 12% AD-11901 −2% AD-11902 40% AD-11903 14%AD-11904 1% AD-11905 33% AD-11906 2% AD-11907 9% AD-11908 5% AD-1190916% AD-11911 37% AD-11912 19% AD-11914 19% AD-11918 1% AD-11919 −1%AD-11925 −5% AD-11926 60% AD-11927 −11% AD-11933 30% AD-11938 6%AD-11939 71% 0.48 AD-11941 47% AD-11942 33% AD-11943 48% AD-11944 51%AD-11945 69% AD-11946 39% AD-11947 39% AD-11948 83% 0.68 AD-11949 41%AD-11950 73% AD-11951 55% AD-11952 81% 1.42 AD-11953 52% AD-11954 55%AD-11955 79% 0.63 AD-11956 37% AD-11957 39% AD-11958 34% AD-11959 36%AD-11960 19% AD-11961 −4% AD-11962 −6% AD-11963 3% AD-11964 13% AD-1196528% AD-11966 −4% AD-11967 15% AD-11968 4% AD-11969 0% AD-11970 −7%AD-11971 3% AD-11972 58% AD-11973 4% AD-11974 38% AD-11975 −11% AD-1197663% 0.21 AD-11977 2% AD-11978 56% AD-11979 5% AD-11980 5% AD-11981 19%AD-11982 65% 0.14 AD-11983 52% AD-11984 50% AD-11985 50% AD-11986 6%AD-11987 −7% AD-11988 58% AD-11989 27% AD-11990 72% 0.24 AD-11991 29%AD-11992 76% 0.94 AD-11993 46% AD-11994 21% AD-11995 −3% AD-11996 53%AD-11997 0% AD-11998 3% AD-11999 19% AD-12000 41% AD-12001 3% AD-1200237% AD-12003 17% AD-12004 5% AD-12005 6% AD-12006 69% AD-12007 81% 0.268AD-12008 35% AD-12009 22% AD-12010 34% AD-12011 10% AD-12012 25%AD-12013 75% 0.60 AD-12014 29% AD-12015 4% AD-12016 2% AD-12017 5%AD-12018 7% AD-12019 79% 0.904 AD-12020 6% AD-12021 0% AD-12022 2%AD-12023 16% AD-12024 87% 0.075 AD-12025 4% AD-12026 5% AD-12027 66%AD-12028 24% AD-12029 −8% AD-12030 46% AD-12031 48% AD-12032 −10%AD-12033 64% 6.74 AD-12034 45% AD-12035 70% 0.11 AD-12036 43% AD-12037AD-12462 35% AD-12463 39% AD-12464 47% AD-12465 34% AD-12466 35%AD-12467 25% AD-12468 −9% AD-12469 −3% AD-12470 50% AD-12471 12%AD-12472 9% AD-12473 52% AD-12474 1% AD-12475 62% 0.13 AD-12476 19%AD-12477 −12% AD-12478 5% AD-12479 21% AD-12480 13% AD-12481 22%AD-12482 65% AD-12483 78% AD-12484 90% 0.023 AD-12485 76% AD-12486 13%AD-12487 60% AD-12488 54% AD-12489 11% AD-12490 72% AD-12491 86% 0.047AD-12492 41% AD-12493 26% AD-12494 12% AD-12495 69% AD-12496 44%AD-12497 2% AD-12498 14% AD-12499 63% AD-12500 86% 0.057 AD-12501 57%AD-12502 88% 0.048 AD-12503 −2% AD-12504 8% AD-12505 29% AD-12506 31%AD-12507 48% AD-12508 47% AD-12509 2% AD-12510 −21% AD-12511 28%AD-12512 43% AD-12513 −22% AD-12514 38% AD-12515 −9% AD-12516 58%AD-12517 18% AD-12518 −8% AD-12519 −5% AD-12520 62% 1.12 AD-12521 −12%AD-12522 53% AD-12523 55% AD-12524 60% AD-12525 32%

TABLE 9 WBC Platelets Lymphocyte # Animal #1 day 0 7.1 328 2 (AD-11570day 3 6.9 308 1.9 treatment) day 5 6.2 394 3.4 Animal #2 day 0 3.6 2991.5 (AD-11570 day 3 10.9 254 1.5 treatment) day 5 12.9 281 1.8 day 821.2 444 4.3 Animal #3 day 0 3.2 218 2.2 (AD-11570 day 3 10.9 202 1.9treatment) day 5 6.4 266 2.4 day 8 18.5 306 3.6 Animal #4 day 0 9.7 3987.3 (untreated) day 3 8.4 448 4.4 day 5 6.2 263 1.9 day 8 2.8 143 1.5

NP Chimeric sequence (SEQ ID NO: 1043):GGTACCCTCGAGGAGGAAGATTAATAATTTTCCTCTCATTGAAATTTATATCGGAATTTAAATTGAAATTGTTACTGTAATCACACCTGGTTTGTTTCAGAGCCACATCACAAAGATAGAGAACAACCTAGGTCTCCGAAGGGAGCAAGGGCATCAGTGTGCTCAGTTGAAAATCCCTTGTCAACACCTAGGTCTTATCACATCACAAGTTCCACCTCAGACTCTGCAGGGTGATCCAACAACCTTAATAGAAACATTATTGTTAAAGGACAGCATTAGTTCACAGTCAAACAAGCAAGATTGAGAATTAACCTTGGTTTTGAACTTGAACACTTAGGGGATTGAAGATTCAACAACCCTAAAGCTTGGGGTAAAACATTGGAAATAGTTAAAAGACAAATTGCTCGGGTTTACCTGAGAGCCTACAACATGGATAAACGGGTGAGAGGTTCATTGGCGCCGAGTCTCACTGAATCTGACATGGATTACCACAAGATCTTGACAGCAGGTCTGTCCGTTCAACAGGGGATTGTTCGGCAAAGAGTCATCCCAGTGTATCAAGTAAACAATCTTGAAGAAATTTGCCAACTTATCATACAGGCCTTTGAAGCAGGTGTTGATTTTCAAGAGAGTGCGGACAGTTTCCTTCTCATGCTTTGTCTTCATCATGCGTACCAGGGAGATTACAAACTTTTCTTGGAAAGTGGCGCAGTCAAGTATTTGGAAGGGCACGGGTTCCGTTTTGAAGTCAAGAAGCGTGATGGAGTGAAGCGCCTTGAGGAATTGCTGCCAGCAGTATCTAGTGGAAAAAACATTAAGAGAACACTTGCTGCCATGCCGGAAGAGGAGACAACTGAAGCTAATGCCGGTCAGTTTCTCTCCTTTGCAAGTCTATTTCTACCCAAACTTGTCGTTGGAGAAAAGGCTTGCCTTGAGAAGGTTCAAAGGCAAATTCAAGTACATGCAGAGCAAGGACTGATACAATATCCAACAGCTTGGCAATCAGTAGGACACATGATGGTGATTTTCCGTTTGATGCGAACAAATTTTCTGATCAAATTTCTCCTAATACACCAAGGGATGCACATGGTTGCCGGGCATGATGCCAACGATGCTGTGATTTCAAATTCAGTGGCTCAAGCTCGTTTTTCAGGCTTATTGATTGTCAAAACAGTACTTGATCATATCCTACAAAAGACAGAACGAGGAGTTCGTCTCCATCCTCTTGCAAGGACCGCCAAGGTAAAAAATGAGGTGAACTCCTTTAAGGCTGCACTCAGCTCCCTGGCCAAGCATGGAGAGTATGCTCCTTTCGCCCGACTTTTGAACCTTTCTGGAGTAAATAATCTTGAGCATGGTCTTTTCCCTCAACTATCGGCAATTGCACTCGGAGTCGCCACAGCACACGGGAGTACCCTCGCAGGAGTAAATGTTGGAGAACAGTATCAACAACTCAGAGAGGCTGCCACTGAGGCTGAGAAGCAACTCCAACAATACGCAGAGTCTCGCGAACTTGACCATCTTGGACTTGATGATCAGGAAAAGAAAATTCTTATGAACTTCCATCAGAAAAAGAACGAAATCAGCTTCCAGCAAACAAACGCTATGGTAACTCTAAGAAAAGAGCGCCTGGCCAAGCTGACAGAAGCTATCACTGCTGCGTCACTGCCCAAAACAAGTGGACATTACGATGATGATGACGACATTCCATTTCCCGGGCCGATCTATGATGACGACAATCCTGGCCATCAAGATGATGATCCGACTGACTCACAGGATACGACCATTCCCGATGGTGTTGTTGACCCGTATGATGGAAGCTACGGCGAATATCCTGACTACGAGGATTCGGCTGAAGGTGCACCAGATGACTTGGTCCTATTCGATCTAGACGAGGACGACGAGGACACTAAGCCAGTGCCTAATAGATCGACCAAGGGTGGACAACAGAAGAACAGTCAAAAGGGCCAGCATATAGAGGGCAGACAGATCCGACCTTGGACGGAGCGAAAAAGGTGCCGGAGTTGCAGAACAATCCACCACGCCAGTGCGCCACTCACGGACAATGACAGAAGAAATGAACCCTCCGGCTCAACCAGCCCTCGCATGCTGACACCAATTAACGAAGAGGCAGACCCACTGGACGATGCCGACGACGAGAGTCTCACATCCCTGCCCTTGGAGTCAGATGATGAAGAGCAGGACAGGGACGGAACTTCCAACCGCACACCCACTGTCGCCCCACCGGCTCCCGTATACAGAGATCACTCTGAAAAGAAAGAACTCCCGCAAGACGAGCAACAAGATCAGGACCACACTCAAGAGGCCAGGAACCAGGACAGTGACAACACCCAGTCAGAACACTCTTTTGAGGAGATGTATCGCCACATTCTAAGATCACAGGGGCCATTTGATGCTGTTTTGTATTATCACCTAATGAGTGATGAGCCTGTAGTTTTCAGTACCAGTGATGGCAAAGAGTACACGTATCCAGACTCCCTTGAAGAGGAATATCCACCATGGCTCACTGAAAAAGAGGCTATGAATGAAGAGAATAGATTTGTTACATTGGATGGTCAACAATTTTATTGGCCGGTGATGAATCACAAGAATAAATTCATGGCAATCCTGCAACATCATCAGTGAATGAGCATGGAACAATGGGATGATTCAACCGACAAATAGCTAACATTAAGTAGTCAAGGAACGAAAACAGGAAGAATTTTTGATGTCTAAGGTGTGAATTATTATCACAATAAAAGTGATTCTTATTTTTGAATTTAAAGCTAGCTTATTATTACTAGCCGTTTTTCAAAGTTCAATTTGAGTCTTAATGCAAATAGGCGTTAAGCCACAGTTATAGCCATAATTGTAACTCAATATTCTAACTAGCGATTTATCTAAATTAAATTACATTATGCTTTTATAACTTACCTACTAGCCTGCCCAACATTTACACGATCGTTTTATAATTAAGAAAAAAGCGGCCGCAGAGCTC GP Chimeric sequence (SEQ ID NO: 1044):GGTACCCTCGAGGATGAAGATTAAGCCGACAGTGAGCGTAATCTTCATCTCTCTTAGATTATTTGTTTTCCAGAGTAGGGGTCGTCAGGTCCTTTTCAATCGTGTAACCAAAATAAACTCCACTAGAAGGATATTGTGGGGCAACAACACAATGGGCGTTCTTAGCCTACTCCAATTGCCTCGTGATCGATTCAAGAGGACATCATTCTTTCTTTGGGTAATTATCCTTTTCCAAAGAACATTTTCCATCCCACTTGGAGTCATCCACAATAGCACATTACAGGTTAGTGAGATTGACCAGCTAGTCTGCAAGGATCATACTGATATGCCATCTGCAACTAAAAGATGGGGCTTCAGGTCCGGTGTCCCACCAAAGGTGGTCAATTATGAAGCTGGTGAATGGGCTGAAAACTGCTACAATCTTGAAATCAAAAAACCGGACGGGAGCGAATGCTTACCCGCAGCGCCAGACGGGATTCGGGGCTTCCCCCGGTGCCGGTATGTGCACAAAGTATCAGGAACGGGACCGTGTGCCGGAGACTTTGCCTTCCATAAAGAGGGTGCTTTCTTCCTGTATGATCGACTTGCTTCCACAGTTATCTACCGAGGAACGACTTTCGCTGAAGGTGTCGTTGCATTTCTGATACTGCCCCAAGCTAAGAAGGACTTCTTCAGCTCACACCCCTTGAGAGAGCCGGTCAATGCAACGGAGGACCCGTCTAGTGGCTACTATTCTACCACAATTAGATATCAGGCTACCGGTTTTGGAACCAATGAGACAGAGTACTTGTTCGAGGTTGACAATTTGACCTACGTCCAACTTGAATCAAGATTCACACCACAGTTTCTGCTCCAGCTGAATGAGACAATATATACAAGTGGGAAAAGGAGCAATACCACGGGAAAACTAATTTGGAAGGTCAACCCCGAAATTGATACAACAATCGGGGAGTGGGCCTTCTGGGAAACTAAAAAAACCTCACTAGAAAAATTCGCAGTGAAGAGTTGTCTTTCACAGTTTTATCGCTCAACGAGACAGACATCAGTGGTCAGAGTCCGGCGCGAACTTCTTCCGGAAGAATCTCCGACCGGGCCACTGAAGACCACAAAATCATGGCTTCAGAAAATTCCTCTGCAATGGTTCAAGTGCACAGTCAAGGAAGGGAAACAACATTGCCGTCTCAGAATTCGACAGAAGGTCGAAGAGCGAGTCCCCAATCCCTCACAACCAAACCAGGTCCGGACAACAGCACCCATAATACACCCGTGTATAAACTTGACATCTCTGAGGCAACTCAAGTTGAACAACATCACCGCAGAACAGACAACGACAGCACAGCCTCCGACACTCCCTCTGCCACGACCGCAGCCGGACCCCCAAAAGCAGAGAACACCAACACGAGCAAGAGCACTGACTTCCTGGACCCCGCCACCACAACAAGTCCCCAAAACCACAGCGAGACCGCTGGCAACAACAACACTCATCACCAAGATACCGGAGAAGAGAGTGCCAGCAGCGGGAAGCTAGGCTTAATTACCAATACTATTGCTGGAGTCGCAGGACTGATCACAGGCGGGAGAAGAACTCGAAGAGAAGCAATTGTCAATGCTCAACCCAAATGCAACCCTAATTTACATTACTGGACTACTCAGGATGAAGGTGCTGCAATCGGACTGGCCTGGATACCATATTTCGGGCCAGCAGCCGAGGGAATTTACATAGAGGGGCTAATGCACAATCAAGATGGTTTAATCTGTGGGTTGAGACAGCTGGCCAACGAGACGACTCAAGCTCTTCAACTGTTCCTGAGAGCCACAACGGAGCTGCGGACATATACCATACTCAACCGTAAGGCAATTGATTTCTTGCTGCAGCGATGGGGCGGCACATGCCACATTCTGGGACCGGACTGCTGTATCGAACCACATGATTGGACCAAGAACATAACAGACAAAATTGATCAGATTATTCATGATTTTGTTGATAAAACCCTTCCGGACCAGGGGGACAATGACAATTGGTGGACAGGATGGAGACAATGGATACCGGCAGGTATTGGAGTTACAGGCGTTATAATTGCAGTTATCGCTTTATTCTGTATATGCAAATTTGTCTTTTAGTTTTTCTTCAGATTGCTTCATGGAAAAGCTCAGCCTCAAATCAATGAAACCAGGATTTAATTATATGGATTACTTGAATCTAAGATTACTTGACAAATGATAATATAATACACTGGAGCTTTAAACATAGCCAATGTGATTCTAACTCCTTTAAACTCACAGTTAATCATAAACAAGGTTTGACATCAATCTAGTTATCTCTTTGAGAATGATAAACTTGATGAAGATTAAGAAAAAGCGGCCGCAGAGCTCThe L gene was generated as 2 fragments (L-ABC, SEQ ID NO:1049; andL-DEFG, SEQ ID NO:1050).

L fragment 1: SEQ ID NO: 1049 (L-ABC):GGCGCGCCTCGAGGAGGAAGATTAAGAAAAACTGCTTATTGGGTCTTTCCGTGTTTTAGATGAAGCAGTTGAAATTCTTCCTCTTGATATTAAATGGCTACCCAACATACACAATACCCAGACGCTAGTTATCATCACCAATTGTATTGGACCAATGTGACCTAGTCACTAGAGCTTGCGGGTTATATTCATCATACTCCCTTAATCCGCAACTACGCAACTGTAAACTCCCGAAACATATCTACCGTTTGAAATACGTGTAACTGTTACCAAGTTCTTGAGTGATGTACCAGTGGCGACATTGCCCATAGATTTCATAGTCCCAGTTCTTCTCAAGGCACTGTCAGGCAATGGATTCTGTCCTGTTGAGCCGCGGTGCCAACAGTTCTAGATGAAATCATTAAGTACACAATGCAAGATGCTCTCTTCTTGAAATATTATCTCAAAAATGTGGGTGCTCAAGAAGACTGTGTTGATGAACACTTTCAAGAGAAAATCTTATCTTCAATTCAGGGCAATGATTTTTACATCAAATGTTTTTCTGGTATGATCTGGCTATTTTAACTCGAAGGGGTAGATTAAATCGAGGAAACTCTAGATCAACATGGTTTGTTCATGATGATTTAATAGACATCTTAGGCTATGGGGACTTGTTTTTTGGAAGATCCCAATTTCAATGTTACCACTGAACACACAAGGAATCCCCCATGCTGCTATGGACTGGTATCAGGCATCAGTATTCAAAGAAGCGGTTCAAGGGCATACACACATTGTTTCTGTTTTACTGCCGACGTCTTGATAATGTGCAAAGATTTAATTACATGTCGATTCAACACAACTCTAATCTCAAAAATAGCAGAGATTGAGGATCCAGTTTGTTCTGATTATCCCAATTTTAAGATTGTGTCTATGCTTACCAGAGCGGAGATTACTTACTCTCCATATTAGGGTCTGATGGGTATAAAATTATTAAGTTCCTCGAACCATTGTGCTTGGCCAAAATTCAATTATGCTCAAAGTACACTGAACGAAAAGGGCGGTTTTAACACAAATGCATTTAGCTGTAAATCACACCCTAGAAGAAATTACAGAAATGCGTGCACTAAAGCCTTCACAGGCTCAAAAGATCCGTGAATTCCATAGAACATTGATAAGGCTGGAGATGACGCCACAACACTTTGTGAGCTATTTTCCATTCAAAAACACTGGGGGCATCCTGTGCTACATAGTGAAACAGCAATCCAAAAAGTTAAAAAACATGCTACGGTGCTAAAAGCATTACGCCCTATAGTGATTTTCGAGACATCTGTGTTTTTAAATATAGTATTGCCAAACATTATTTTGATAGTCAAGGATCTTGGTACAGTGTTACTTCAGACCGATGTTTAACGCCGGGATTGAATTCTTATATCAAAAGAAATCAATTCCCTCCGTTGCAATGATTAAAGAACTACTATGGGAATTTTACCACCTTGACCACCCTCCACTTTTCTCAACCAAAATTATTAGTGACTTAAGTATTTTTATAAAAGACAGAGCTACCGCAGTAGAAAGGACATGCTGGGATGAGTATTCGAGCCTAATGTTCTAGGATATAATCCACCTCACAAATTTAGTACTAAACGTGTACCGGAACAATTTTTAGAGCAAGAAAACTTTTCTATTGAGAATGTTCTTTCATACGCCCAAGAACTTAGGTTCTACTACCACAATATCGGAACTTTTCTTTCTCATTGAAAGAGAAAGAGTTGAATGTAGGTAGAACCTTCGGAAAATTGCCTTATCCGACTCGCAATGTTCAAACACTTTGTGAAGCTCTGTTAGCTGATGTCTTGCTAAAGCATTTCCTAGCAATATGATGGTAGTTACGGAACGTGAGCAAAAAGAAAGCTTATTGCATCAAGCATCATGGCACCACACAAGTGATGATTTTGGTGAACATGCCACAGTTAGAGGGAGTACTTTGTAACTGATTTAGAGAAATACAATCTTGCATTTAGATATGAGTTTACAGCACCTTTTATAGAATATTGCAACCGTTGCTATGGTGTTAAGAATGTTTTTAATTGGATGCATTATACAATCCCACAGTTTATATGCATGTCAGTGATTATTATAATCCACCACATAACCTCACACTGGAGAATCGAGACAACCCCCCCGAAGGGCCTAGTTCATACAGGGGTCATATGGGAGGGATTGAAGGACTGCAACAAAAACTCTGACAAGTATTTCATGTGCTCAAATTTCTTTAGTTGAAATTAAGACTGGTTTTAAGTTACGCTCAGCTGTGATGGGTGACAATCAGTGCATTACTGTTTTATCAGTCTTCCCCTTAGAGACTGACGCAGACGGCAGGAACAGAGCGCCGAAGACAATGCAGCGAGGGTGGCCGCCAGCCTAGCAAAAGTTACAAGTGCCTGTGGAATCTTTTTAAAACCTGATGAGACTTTCGTACACTCAGGTTTTATCTATTTTGGAAAAAACAATATTTGAATGGGGTCCAATTGCCTCAGTCCCTTAAAACGGCTACAAGAATGGCACCATTGTCTGATGCAATTTTTGATGATCTTCAAGGGACCCTGGCTAGTATAGGCACTGCTTTTGAGCGATCAACTCCGAAACTAGACATATCTTTCCTTGCAGGATAACCGCAGCTTTCCATACGTTTTTTTCGGTGAGAATCTTGCAATATCATCATCTCGGGTTCAATAAAGGTTTTGACCTTGGACAGTTAACACTCGGCAACCTCTGGATTTCGGAACAATATCATTGGCACTAGCGGTACCGCAGGTGCTTGGAGGGTTATCCTTCTTGAATCCTGAGAAATGTTTCTACCGGAATCTAGGAGATCCAGTTACCTCAGGCTTATTCCAGTAAAAACTTATCTCCGAATAGAGACCTATTGAGCTCCACCGCGGTGGCGGCCGCTCTAGCCCGGGCGGATCCCCCGGGCTGCAGGAATTCGATATCAAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGTACCTTACATCGCGTTAATTAACTAGTGGATCGATCCCCAATTCGL fragment 2: SEQ ID NO: 1050 (L-DEFG):CGCGACGTAATACGACTCACTATAGGGCGAATTGGGCGCGCCCCGTCTCAGAATGATTGAGATGGATGATTTATTCTTACCTTTAATTGCGAAGAACCCTGGGAATTGTAGCGCAATTGACTTTGTGTAAATCCTAGCGGATTAAATGTCCCTGGGTCGCAAGACTTAACTTCATTTCTGCGCCAGATTGTACGCAGGACCATCACCCTAAGTGCGAAAAACAAACTTATTAATACCTTATTTCATGCGTCAGCTGACTCGAAGACGAAATGGTTTGTAAATGGCTATTATCATCAACTCCTGTTATGAGTCGTTTTGCGGCCGATATCTTTTCACGCACGCCGAGCGGGAAGCGATTGCAAATTCTAGGATACCTGGAAGGAACACGCCATTATTAGCCTCTAAGATCATCAACAATAATACAGAGACACCGGTTTTGGACAGACTGAGGAAAATAACATTGCAAAGGTGGAGCCTATGGTTTAGTTATCTTGATCATTGTGATAATATCCTGGCGGAGCTTTAACCCAAATAACTTGCACAGTTGATTTAGCACAGATTCTGAGGGAATATTCATGGGCTCATATTTTAGAGGGAAGACCTCTTATTGGAGCCACACTCCCATGTATGATTGAGCAATTCAAAGTGTTTGGCTGAAACCCTACGAACAATGTCCGCAGTGTTCAAATGCAAAGCAACCAGGTGGGAAACCATTCGTGTCAGTGGCAGTCAAGAAACATATTGTTAGTGCATGGCCGAACGCATCCCGAATAAGCTGGACTTCGGGGATGGAATCCCATACATTGGATCAAGGACAGAAGATAAGATAGGTCAGCCCGCTATTAAGCCGAGGTGTCCTTCCGCAGCCTTAAGAGAGGCCATTGAATTGGCGTCCCGTTTAACATGGGTAACTAAGGCAGTTCGAACAGTGACTTGCTAATAAAACCATTTTTGGAAGCACGAGTAAATTTAAGTGTTCAAGAAATACTTCAAATGACCCCTTCACATTACTCAGGAAATATTGTGCATCGGTATAACGATCAAACAGTCCTCATTCTTTCATGGCCAATCGTATGAGTAATTCAGCAACGCGCTTGATGGTATCTACAAACACTTTAGGTGAGTTTTCAGGAGGTGGCCAGTCTGCACGCGACAGCAATATTATTTTCCAGAATTTATAAATTATGCAGTTGCACTGTTCGATATTAAATTTAGAAACACTGAGGCTACAGATATCCAATATAATCGTGCTCACCTTCATCTAACTAAGTGTTGCACCCGGGAAGTACCAGCTCAGTATTTAACAACACAACCACGCTAAATCTAGATTTAACAAGATACCGAGAAAACGAATTGATTTATGACAGTAATCCTCTAAAAGGAGGACTCAATTGCAACTTATCGATTGACAGTCCTTTTTTCCAAGGTAAACGGCTGACATTATAGAAGATGATCTTATTCGACTGCCTCACTTATCTGGATGGGAGCTAGCCAAGACCATCATGCAATCAATTATTTCAGATAGCAACAATTCATCTACAGACCCAATTAGCAGTGGAGAAACAAGACATTCACTACCCATTTCTTAACTTATCCCAAGATAGGACTTCTGTACAGTTTTGGGGCCTTTGTAAGTTATTATCTTGGCAATACAATTCTTTGCACGAAAAAGATCGGACTTGACAATTTTTTATATTACTAACTACTCAAATTCATAATCTACCACATCGCTCATTGCGAATACTTAAGCCAACATTCAAACATGCAAGCGTTATGTCACGGTTAATGAGTATTGATCCTCATTTTTCTATTTACATAGGCGGTGCTGCAGTGACAGAGGACTCTCAGATGCGGCCAGGTTATTTTTGAGAACGTCCATTTCATCTTTTCTTACATTTGTAAAAGAATGGATAATTAATCGCGGAACAATTGTCCCTTTATGGATAGTATATCCGCTAGAGGTCAAAACCCAACACCTGTGAATAATTTTCTCTATCAGATCGTAGAACTGCTGGTGCATGATTCATCAAGACAACAGGCTTTTAAAACTACCATAAGTGATCATGTACATCCTCACGACAATCTTGTTTACCATGTAAGAGTACAGCCAGCAATTTCTTCCATGCATCATTGGCGTACTGGAGGAGCAGACACAGAAACAGCAACCGAAAATACTTGGCAAGAGACTCTTCAACTGGATCAAGCACAAACAACAGTGATGGTATATTGAGAGAAGTCAAGAACAAACCACCAGAGATCCACATGATGGCACTGAACGGAATCTAGTCCTACAAATGAGCCATGAAATAAAAAGAACGACAATTCCACAAGAAAACACGCACCAGGGTCCGTCGTCCAGTCCTTTCTAAGTGACTCTGCTTGTGGTACAGCAAATCCAAAACTAAATTTCGATCGATCGAGACACAATGTGAAATTTCAGGATCATAACTCGGCATCCAAGAGGGAAGGTCATCAAATAATCTCAACCGTCTAGTCCTACCTTTCTTTACATTATCTCAAGGGACACGCCAATTAACGTCATCCAATGAGTCACAAACCCAAGACGAGATATCAAAGTACTTACGGCAATTGAGATCCGTCATTGATACTACCATAATTGTCGCTTCACCGGTATAGTCTCGTCCATGCATTACAAACTTGATGAGGTCCTTTGGGAAATAGAGAGTTTCAAGTCGGCTGTGACGCTAGCAGAGGGAGAAGGTGCTGGTGCCTTACTATTGATTCAAAATACGGCGTTAAGAAGTTATTTTTCAACACGCTAGCTACTGAGTCCAGTATAGAGTCAGAAATAGTATCAGGAATGACTACTCCTAGGATGCTTCTACCTGTTATGTCAAAATTCCATAATGACCAAATTAGATTATTCTTAACAACTCAGCAAGCCAAATAACAGACATAACAAATCCTACTTGGTTTAAAGACCAAAGAGCAAGGCTACCTAAGCAAGTCGAGGTTATAACCATGGATGCAGAGACAACAGAGAATATAACAGATCGAAATTGTACGAAGCTGTATATAAATTGATCTTACACCATATTGATCCTAGCGTATTGAAAGCAGTGGTCCTTAAAGTCTTTCTAAGTGATACTGAGGGTATGTTATGGCTAAATGATAATTTACCCCGTTTTTTGCCACTGGTTATTTAATTAAGCCAATAACGTCAAGTGCTAGATCTAGTGAGTGGTATCTTTGTCTGACGAACTTCTTATCAACTACACGTAAGATGCCACACCAAAACCATCTCAGTTGTAACAGGTAATACTTACGGCATTGCAACTGCAAATTCAACGAAGCCCATACTGGCTAAGTCATTTAACTCAGTATGCTGACTGTGAGTTACATTTAAGTTATATCCGCCTTGGTTTTCCATCATTAGAGAAATACTATACCACAGGTATAACCTCGTCGATTCAAAAAGAGGTCCACTAGTCTCTATCACTCAGCACTTAGCACATCTTAGAGCAGAGATTCGAGAATTAACTAATGATTATAATCAACAGCGACAAAGTCGACCCAGACTTATCATTTTATTCGTACTGCAAAAGGACGGATAACTAAACTAGTCAATGATTATTTAAAATTCTTTCTTATTGTGCAAGCATTAAAACATAATGGGACATGGCAAGCTGAGTTTAAGAAATTACAGAGTTGATTAGTGTGTGCAATAGGTTCTACCATATTAGAGATTGCAATTGTGAAGAACGTTTCTTAGTTCAAACCTTATATTTACATAGAATGCAGGATTCTGAAGTTAAGCTTATCGAAAGGCTGACAGGCTTCTGAGTTTATTTCCGGATGGTCTCTACAGGTTTGATTGAATTACCGTGCATAGTATCCTGATACTTGCAAAGGTTGGTTATTAACATACAGATTATAAAAAAGCGGCCGCAGAGCTCCAGCGGTGGGGCCGCCGGCGTCTAGCCCGGGCGGATCCCTGCAGGAATTCGATATCAAGCTTATCGATACCGTCGACCTCGAGGGCCCATGCAGGCCGGCCAGGTACCTTAGTTAATTAACAGCTTTTGTTCCCTTTAGTAGGGTTAATTGACGCGCTC VP24 (SEQ ID NO: 1045):GGCGCGCCTCGAGGATGAAGATTAATGCGGAGGTCTGATAAGAATAAACCTTATTATTCAGATTAGGCCCCAAGAGGCATTCTTCATCTCCTTTTAGCAAAGTACTATTTCAGGGTAGTCCAATTAGTGGCACGTCTTTTAGCTGTATATCAGTCGCCCCTAGGCTAGGGTTTATAGTTGTCTCTAAGCTAAATTGGTCTTGAACTAGTCTACTCGCAGAATCCTACCGGGAATAGACTAATTGAACTTAGCCGTTTAAAATTTAGTGCATAAATCTGGGCTAACACCACCAGGTCAACTCCATTGGCTGAAAAGAAGCTTACCTACAACGAACATCACTTTGAGCGCCCTCACAATTAAAAAATAGGAACGTCGTTCCAACAATCGAGCGCAAGGTTTCAATATTATACGGGTCCATTAATTTCAACAAAATATTGATACTCCAGACACCAAGCAAGACCTGAGAAAAAACCATGGCTAAAGCTACGGGACGATACAATCTAATATCGCCCAAAAAGGACCTGGAGAAAGGGGTTGTCTTAAGCGACCTCTGTAACTTCTTAGTTAGCCAAACTATTCAGGGGTGGAAGGTTTATTGGGCTGGTATTGAGTTTGATGTGAACCAAAAGGGTATTACCCTATTGCATAGACTGAAAACTAATGACTTTGCCCCTGCATGGTCAATGACAAGGAATCTCTTTCCTCATTTATTTCAAAATCCGAATTCCACAATTGAATCACCGCTGTGGGCATTGAGAGTCATCCTTGCAGCAGGGATACAGGACCAGCTGATTGACCAGTCTTTGATTGAACCCTTAGCAGGAGCCCTTGGTCTGATCTCTGATTGGCTGCTAACAACCAACACTAACCATTTCAACATGCGAACACAACGTGTCAAGGAACAATTGAGCCTAAAAATGCTGTCGTTGATTCGATCCAATATTCTCAAGTTTATTAACAAATTGGATGCTCTACATGTCGTGAACTACAACGGATTGTTGAGCAGTATTGAAATTGGAACTCAAAATCATACAATCATCATAACTCGGACTAATATGGGTTATCTTGTCGAGCTCCAAGAACCCGACAAATCTGCGATGGATATACGACACCCTGGGCCGGCGAAATTTTCCTTACTACATGAATCGACACTTAAAGCATTTACACAAGGATCCTCGACACGAATGCAAAGTTTGATTCTTGAATTTAATAGCTCTCTTGCTATCTAACTAAGGTAGAAAAAATTGTACGATAGGGCTAACATTATGCTGACTCAATAGTTATCTTGACATCTCTGCTTTCATAATCAGATATATAAGCATAATAAATAAATACTCATATTTCTTGATAATTTGTTTAACCACAGATAAATCCTCACTGTAAGCCAGCTTCCAAGTTGACACCCTTACAAAAACCAGGACTCAGAATCCCTCAAACAAGAGATTCCAAGACAACATCATAGAATTGCTTTATTATATGAATAAGCATTTTATCACCAGAAATCCTATATACTAAATGGTTAATTGTAACTGAACCCGCAGGTCACATGTGTTAGGTTTCACAGATTCTATATATTACTAACTCTAGAGCCCAAATTAACACGGTATAAGTAGATTAAGAAAAAAGCCTGAGGAAGATTAAGAAAAAGCGGCCGCATTAATTAA VP30 (SEQ ID NO: 1046):GGCGCGCCTCGAGGATGAAGATTAAGAAAAAGGTAATCTTTCGATTATCTTTAATCTTCATCCTTGATTCTACAATCATGACAGTTGTCTTTAGTGACAAGGGAAAGAAGCCTTTTTATTAAGTTGTAATAATCAGATCTGCGAACCGGTAGAGTTTAGTTGCAACCTAACACACATAAAGCATTGGTCAAAAAGTCAATAGAAATTTAAACAGTGAGTGGAGACAACTTTTAAATGGAAGCTTCGTGAGCGCGGGAGATCAAGGAATTCACGTGCCGACCAGCAAGGGATGGACACGACCACCATGTTCGAGCACGATCATCATCCAGAGAGAATTATCGAGGTGAGTACCGTCAATCAAGGAGCGCCTCACAAGTGCGCGTTCCTACTGTATTTCATAAGAAGAGAGTTGAACCATTAACAGTTCCTCCAGCACCTAAAGACATATGTCCGACCTTGAAAAAAGGATTTTTGTGTGACAGTAGTTTTTGCAAAAAAGATCACCAGCTTGAAAGCCTAACCGACCGGGAATTACTCCTACTAATCGCCCGTAAGACTTGTGGATCAGTTGATTCATCGCTTAATATAACTGCACCCAAGGACTCGCGCTTAGCAAATCCAACGGCTGATGATTTCCAGCAAGAGGAAGGTCCAAAAAATTACTAGTCGAGACTGCTCAAGACGGCAGAACACTGGGCGAGACAAGACATCAGAACCATAGAGGATTCAAAATTAAGAGCATTGTTGACTCTATGTGCTGTGATGACGAGGAAATTCTCAAAATCCCAGCTGAGTCTTTTATGTGAGACACACCTAAGGCGCGAGGGGCTTGGGCAAGATCAGGCAGAACCCGTTCTCGAAGTATATCAACGATTACACAGTGATAAAGGAGGCAGTTTTGAAGCTGCACTATGGCAACAATGGGACCGACAATCCCTAATTATGTTTATCACTGCATTCTTGAATATTGCTCTCCAGTTACCGTGTGAAAGTTCTGCTGTCGTTGTTTCAGGCCTACGCTTACTTGCCCCCCCAAGCGTTAATGAAGAAGCTTCAACCAACCCGGGGACATGCTCATGGTCTGATGAGGGTACCCCTTAATAAGGCTGACTAAAACACTATATAACCTTCTACTTGATCACAATACTCCGTATACCTATCATCATATATTTAATCAAGACGATATCCTTTAAAACTTATTCAGTACTATAATCACTCTCGTTTCAAATTAATAAGATGTGCATGATTGCCCTAATATATGAAGAGGTATGATACAACCCTAACAGTGATCAAAGAAAATCATAATCTCGTATCGCTCGTAATATAACCTGCCAAGCATACTCCCTAGAAGCGTTGAATCTTGTACACAAATAATGTTTTACTCTACAGGAGGTAGCAACGATCCATCCCATCAAAAAATAAGTATTTCATGACTTACTAATGATCTCTTAAAATATTAAGAAAAAGCGGCCGCATTAATTAAVP35 (SEQ ID NO: 1047):GGTACCGCGATCGCGATGAAGATTAAAACCTTCATCATCCTTACGTCAATTGAATTCTCTAGCACTCGAAGCTTATTGTCTTCAATGTAAAAGAAAAGCTGGTCTAACAAGATGACAACTAGAACAAAGGGCAGGGGCCATACTGCGGCCACGACTCAAAACGACAGAATGCCAGGCCCTGAGCTTTCGGGCTGGATCTCTGAGCAGCTAATGACCGGCAAAATACCGCTAACCGACATCTTCTGTGATATTGAGAACAATCCAGGATTATGCTACGCATCCCAAATGCAACAAACGAAGCCAAACCCGAAGACGCGCAACAGTCAAACCCAAACGGACCCAATTTGCAATCATAGTTTTGAGGAGGTAGTACAAACATTGGCTTCATTGGCTACAGCTGTGCGTCGGCAAACCATCGCATCAGAATCATTAGAACAACGCATTACGAGTCTTGAGAATGGTCTAAAGCCAGTTTATGATATGGCAAAAACAATATCATCCCTGAATCGCAGCTGTGCTGAGATGGTTGCAAAATATGATCTTCTGGTGATGACAACCGGTCGGGCAACAGCAACCGCTGCGGCAACTGAGGCTTATTGGGCCGAACATGGTCAACCACCACCAGGCCCATCATTGTACGAGGATGGTGCGATTCGGGGTAAATTGAAAGATCCGAACGGGACCGTCCCTCAAAGTGTTAGGGAGGCATTCAACAATCTAAACAGTACCACTTCACTAACTGAGGAAAATTTCGGGCGACCTTACATTTCGGCAAAGGATTTGAGAAACATTATGTATGATCACTTGCCTGGTTTTGGAACTGCTTTCCACCAATTAGTACAAGTGATTTGTAAATTGGGAAAAGATAGCAACTCATTGGACATCATTCATGCTGAGTTCCAGGCCAGCCTGGCTGAAGGAGACTCTCCTCAATGTGCCCTAATTCAAATTACAAAAAGAGTTCCAATCTTCCAAGATGCTGCTCCACCTGTCATCCACATCCGCTCTCGAGGTGACATTCCCCGAGCTTGCCAGAAAAGCTTGCGTCCAGTCCCACCATCGCCCAAGATTGATCGAGGTTGGGTATGTGTTTTTCAGCTTCAAGATGGTAAAACACTTGGACTCAAAATTTGAGCCAATGTAAGCTCATTTTGCGATGGGCGAATAATAGCAGAGGCTTCAACTGCTGAACTATAGGGTACGTTACATTAATGATACACTTGTGAGTATCAGCCCTGGATAATATAAGTCAATCCTAATCAATTGATAATATTGTTCATATCTCGCTAGCAGCTTAAAATATAAATGTAATAGGAGCTATATCTCTGACAGTATTATAATCAATTGTTATTAAGTAACCCAAACCAAAAGTGATGAAGATTAAGAAAAAGCGGCCGCAGAGCTC VP40 (SEQ ID NO: 1048):GGTACCTCGAGGATGAAGATTAAGAAAAACCTACCTCGGCTGAGAGAGTGTTTTTTCATTAACCTTCATCTTGTAAACGTTGAGCAAAATTGTTAAAAATATGAGGCGGGTTATATTGCCTACTGCTCCTCCTGAATATATGGAGGCCATATACCCTGTCAGGTCAAATTCAACAATTGCTAGAGGTGGCAACAGCAATACAGGCTTCCTGACACCGGAGTCAGTCAATGGGGACACTCCATCGAATCCACTCAGGCCAATTGCCGATGACACCATCGACCATGCCAGCCACACACCAGGCAGTGTGTCATCAGCATTCATCCTTGAAGCTATGGTGAATGTCATATCGGGCCCCAAAGTGCTAATGAAGCAAATCCCTATTTGGTTGCCTCTAGGTGTCGCTGATCAAAAGACCTACAGCTTTGACTCAACTACGGCCGCAATTATGCTCGCATCTTATACGATCACCCATTTCGGCAAGGCAACCAACCCCCTCGTTAGAGTGAATCGACTGGGTCCTGGAATCCCGGATCATCCCCTCAGGCTCCTGCGAATTGGAAACCAGGCTTTCCTCCAGGAGTTCGTTCTTCCGCCAGTCCAACTACCCCAGTATTTCACCTTTGATTTGACAGCACTCAAACTGATCACCCAACCACTGCCTGCTGCAACATGGACCGATGACACTCCAACAGGATCAAATGGAGCGTTGCGTCCAGGAATTTCATTTCATCCAAAACTTCGCCCCATTCTTTTACCCAACAAAAGTGGGAAGAAGGGGAACAGTGCCGATCTAACATCTCCGGAGAAAATCCAAGCAATAATGACTTCACTCCAGGACTTTAAGATCGTGCCAATTGATCCAGCCAAGAGTATCATTGGGATCGAGGTGCCAGAAACTCTGGTCCACAAGCTGACCGGTAAGAAGGTGACTTCTAAAAATGGACAACCAATCATCCCTGTTCTTTTGCCAAAGTACATTGGGTTGGACCCGGTGGCTCCAGGAGACCTCACCATGGTAATCACACAGGATTGTGACACGTGTCATTCTCCTGCAAGTCTTCCAGCTGTGATTGAGAAGTAATTGCAATAATTGACTCAGATCCAGTTTTATAGAATCTTCTCAGGGATAGCAACTCAATCGACTTTTAGGACCGTCCATTAGAGGAGACACTTTTAATTGAAAAATGTACTAATCGGGTCAAGGACCATTGTCTTTTTTCTCTCCTAAATGTAGAACTTAACAAAAGACTCATAATATACTTGTTTTTAAAGGATTGATTGATGAAAGAACATGCATAAGCGATCCATACTTCGCCCTACTATAATCAATACGGTGATTCAAATGTTAATCTTTCTCATTGCACATACTTTTTGCCCTTATCCTCAAATTGCCTGCATGCTTACATCTGAGGATAGCCAGTGTGACTTGGATTGGAAATGTGGAGAAAAAATCGGGACCCATTTCTAGGTTGTTCACAATCCAAGTACAGACATTGCCCTTCTAATTAAGAAAAAAGCGGCCGCAGAGCTC

Other embodiments are in the claims.

We claim:
 1. A double-stranded ribonucleic acid (dsRNA) for inhibitingthe expression of a gene in the Ebola virus in a cell, wherein saiddsRNA comprises at least two sequences that are complementary to eachother and wherein a sense strand and an antisense strand form a duplexstructure less than 30 nucleotides in length, and wherein the sensestrand is complementary to SEQ ID NO:
 579. 2. The dsRNA of claim 1,wherein said dsRNA comprises at least one modified nucleotide.
 3. ThedsRNA of claim 2 wherein said modified nucleotide is selected from thegroup consisting of: a 2′-O-methyl modified nucleotide, a nucleotidecomprising a 5′-phosphorothioate group, and a terminal nucleotide linkedto a cholesteryl derivative or dodecanoic acid bisdecylamide group. 4.The dsRNA of claim 2, wherein said modified nucleotide is selected fromthe group consisting of: a 2′-deoxy-2′-fluoro modified nucleotide, a2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide,2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholinonucleotide, a phosphoramidate, and a non-natural base comprisingnucleotide.
 5. A cell comprising the dsRNA of claim
 1. 6. Apharmaceutical composition for inhibiting the expression of a gene froman Ebola virus in an organism, comprising a dsRNA of claim 1 and apharmaceutically acceptable carrier.
 7. The pharmaceutical compositionof claim 6, wherein said sense strand of said dsRNA consists of thesequence of SEQ ID NO: 579, and said antisense strand consists of thesequence of SEQ ID NO:
 580. 8. A method for inhibiting the expression ofa gene from an Ebola virus in a cell, the method comprising: (a)introducing into the cell a double-stranded ribonucleic acid (dsRNA) ofclaim 1; and (b) maintaining the cell produced in step (a) for a timesufficient to obtain degradation of the mRNA transcript of a gene fromthe Ebola virus, thereby inhibiting expression of a gene from the Ebolavirus in the cell.
 9. A method of treating a pathological processmediated by Ebola expression comprising administering to a patient atherapeutically effective amount of the dsRNA of claim
 1. 10. A vectorcomprising a regulatory sequence operably linked to a nucleotidesequence that encodes the dsRNA of claim
 1. 11. A cell comprising thevector of claim
 10. 12. The dsRNA of claim 1, wherein the dsRNA has asense strand consisting of the sequence of SEQ ID NO: 579, and antisensestrand consisting of the sequence of SEQ ID NO:
 580. 13. The dsRNA ofclaim 1, wherein said dsRNA, upon contact with a cell infected withEbola virus, inhibits expression of a gene from the virus by at least40%.
 14. The dsRNA of claim 1, wherein said duplex structure is 15-30nucleotides in length.
 15. The dsRNA of claim 1, wherein the dsRNA has asense strand comprising the sequence of SEQ ID NO:579, and an antisensestrand comprising the sequence of SEQ ID NO:580.
 16. The pharmaceuticalcomposition of claim 6, wherein the dsRNA has a sense strand comprisingthe sequence of SEQ ID NO:579, and an antisense strand comprising thesequence of SEQ ID NO:580.
 17. The dsRNA of claim 1, wherein said duplexstructure is 19-21 nucleotides in length.